Doxepin analogs and methods of use thereof

ABSTRACT

The invention relates to novel antihistamines and methods of modulating sleep by administering a doxepin analog or a pharmaceutically effective salt thereof.

RELATED APPLICATIONS

This application claims priority under 35 U.S.C. 119(e) to U.S.provisional patent application Ser. No. 60/528,393, filed Dec. 10, 2003,the entire contents of which are incorporated by reference herein.

FIELD OF THE INVENTION

The invention relates to methods for treating sleep disorders andcompositions useful in such methods.

BACKGROUND OF THE INVENTION

Difficulty falling asleep or remaining asleep is a significant medicalissue that arises for a variety of reasons. Sometimes, these problemsarise from endogenous conditions such as sleep apnea or insomnia. Othertimes, these problems arise from exogenous stresses such as thedisruptive effect of shift work schedules and “jet lag.” Whether causedby an endogenous or exogenous source, difficulty falling asleep orremaining asleep can result in problem sleepiness, which impairs thehealth, quality of life and safety of those affected.

Existing pharmaceutical treatments for inducing sleep include sedativesor hypnotics such as benzodiazepine and barbiturate derivatives. Thesetreatments have numerous drawbacks, including rebound insomnia, delayedonset of desired sedative effects, persistence of sedative effects afterthe desired sleep period, and side effects due to nonspecific activitysuch as psychomotor and memory deficits, myorelaxation, and disturbedsleep architecture, including REM sleep inhibition. Additionally,sedatives and hypnotics can be habit forming, can lose theireffectiveness after extended use and may be metabolized more slowly bysome people.

Consequently, physicians often recommend or prescribe antihistamines asa milder treatment for sleep disorders when hypnotics are lessappropriate. However, many antihistamines still have a number of sideeffects. Other issues include prolongation of the QT interval in asubject's electrocardiogram, as well as central nervous system (CNS)side effects such as decreased muscle tone, drooping eyelids anddrowsiness. Finally, such compounds can bind to muscarinic receptors,which leads to anti-cholinergic side effects such as blurred vision, drymouth, constipation, urinary problems, dizziness and anxiety.

As a result, there is a need for sleep-related treatments that areantihistamines with reduced side effects. Additionally, while knownsleep-inducing compounds are effective for treating sleep-onsetinsomnia, i.e., a subject's difficulty in falling asleep, there are nodrugs currently indicated for treating sleep maintenance insomnia, ie.,maintaining a subject's sleep throughout a normal sleep period afterfalling asleep. Therefore, there is also a need for improvedpharmaceutical treatments for maintaining sleep in subjects in need ofsuch treatment.

SUMMARY OF THE INVENTION

The present invention relates to doxepin analogs compounds and their useto modulate sleep.

In one aspect, the invention provides a method of modulating sleep in asubject by administering a therapeutically effective amount of acompound having the formula of Formula I:

or a pharmaceutically effective salt thereof, wherein m n, o, p, q are,individually, 0-6; X and Y are, individually, absent, O, S, C(O), SO orSO₂; R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are, independently selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chainalkyl, C₃-C₆ branched alkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃,OCF₃, CH₂OCH₃, CH₂CH₂OCH₃, CH₂OCH₂CH₃, and C₁-C₆ hydroxyalkyl; anyhydrogen in the CH₂ groups in the linker is optionally substituted withH, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chain alkyl, C₃-C₆ branchedalkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃, OCF₃, CH₂OCH₃,CH₂CH₂OCH₃, CH₂OCH₂CH₃, or C₁-C₆ hydroxyalkyl; R₉, R₁₀, R₁₁, and R₁₂are, independently, H, C₁-C₅ straight chain alkyl, C₂-C₆ branched alkyl,R₉ and R₁₀ together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to 7, or R₁₁ and R₁₂ togetherwith the carbon to which they are attached, are connected to form aspiro ring of size 3 to 7; or substituents on two different atoms areconnected to form a ring of size 3 to 7; R₁₃ and R₁₄ are, independently,selected from the group consisting of H, F, Cl, Br, CH₃, C₁-C₆ straightchain alkyl, and C₂-C₆ branched alkyl; and Z is selected from the groupconsisting of CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl,CONHS(O)₂-Heteroaryl, SO₃H, SO₂H, S(O)₂NHCO-alkyl, S(O)₂NHCO-aryl,S(O)NHCO-alkyl, S(O)NHCO-aryl, P(O)(OH)₂, P(O)OH,

the compound has one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 500 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, M3, D1, D2, D3, α1and α2 that is more than 10 times greater than the K_(i) with regard tothe H1 receptor; (iii) a nonREM peak time value that is greater than 55%nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) and administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.

In one embodiment, the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 150 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, and M3, that is greater than 10 μM; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 17 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 5 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 6 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity or motor tonerelative to the normal effects of sleep.

The methods of the invention are used to treat a variety of subjects,including, for example, humans, companion animals, farm animals,laboratory animals and wild animals.

In one embodiment, the compound used in this method of modulating sleepis 47a, 47b, 47c, 47d, 49a, 49b, 49c, 49d, 10a, 10b, 10c, 10d, 37, 38a,38b, 39a or 39b.

In one embodiment, the R₉ and R₁₀ and the carbon they are attached toare absent. In one embodiment, R₁₁ and R₁₂, together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7. For example, R₁₁ and R₁₂ together with the carbon to which they areattached, are connected to form a spiro 3-membered cyclopropyl ring.

In one embodiment, Z is CO₂H or

In one embodiment where Z is COOH, at least one of R₁-R₈, R₁₃-R₁₄ and atleast one of R₉-R₁₀, R₁₁-R₁₂ are not hydrogen.

In one embodiment, R₁, R₃-R₈ and R₁₃-R₁₄ are each hydrogen and R₂ is notH, CH₃, CF₃, Cl or Br. In another embodiment, R₁, R₃-R₅, R₇-R₈ andR₁₃-R₁₄ are each hydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃, CF₃, Cl orBr. In one embodiment, R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ are each hydrogen,R₆ is CH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In anotherembodiment, each of R₁, R₃-R₈ and R₁₃-R₁₄ is H and R₂ is F. In oneembodiment, each of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ is H, R₆ is OCH₃, andR₂ is OCH₃. In another embodiment, each of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄is H, R₆ is F, and R₂ is OCH₃.

In one embodiment, at least three of R₁-R₈ are not hydrogen. In oneembodiment, at least one of R₁₃-R₁₄ is not hydrogen. In one embodiment,R₃ is not hydrogen. In one embodiment, R₇ is not hydrogen.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl. In one embodiment, R₁₁ and R₁₂ are each ethyl.In another embodiment, R₁₁ and R₁₂ are each ethyl.

In one embodiment, R₁₁ and R₁₂ and the carbon to which they are attachedare connected to form a spiro ring of size 3-7. For example, in oneembodiment, R₁₁ and R₁₂ and the carbon to which they are attached areconnected to form a three-membered spiro (cyclopropyl) ring.

In one aspect, the compounds of the invention are used to modulatesleep, e.g., by decreasing the time to sleep onset, increasing theaverage sleep bout length, and/or increasing the maximum sleep boutlength. In another aspect, the doxepin analogs of the invention are usedto treat a sleep disorder. For example, the doxepin analogs of theinvention are used to treat circadian rhythm abnormality, insomnia,parasomnia, sleep apnea syndrome, narcolepsy and/or hypersomnia.

In one embodiment, the doxepin analogs of the invention are used in thetreatment of a circadian rhythm abnormality, such as, for example, jetlag, shift-work disorders, delayed sleep phase syndrome, advanced sleepphase syndrome and non-24 hour sleep-wake disorder.

In another embodiment, the doxepin analogs are used in the treatment ofinsomnia, including, for example, extrinsic insomnia, psychophysiologicinsomnia, altitude insomnia, restless leg syndrome, periodic limbmovement disorder, medication-dependent insomnia, drug-dependentinsomnia, alcohol-dependent insomnia and insomnia associated with mentaldisorders.

In one embodiment, the doxepin analogs of the invention are used totreat a parasomnia disorder, such as, e.g., somnambulism, pavornocturnus, REM sleep behavior disorder, sleep bruxism and sleepenuresis.

In another embodiment, the doxepin analogs are used to treat a sleepapnea disorder, such as, for example, central sleep apnea, obstructivesleep apnea and mixed sleep apnea.

Pharmaceutical compositions that include a compound of Formula I or apharmaceutically acceptable salt thereof are used in the methods ofmodulating sleep. In one embodiment, the compound of Formula I orpharmaceutically acceptable salt thereof is co-administered with one ormore additional therapies.

In another aspect, the present invention provides a method of modulatingsleep in a subject by administering a therapeutically effective amountof a compound having the formula of Formula II:

or a pharmaceutically effective salt thereof, wherein m, n, and o are,individually, 0-6; X is absent, O, S, C(O), SO or SO₂; R₁, R₂, R₃, R₆,and R₇ are, independently selected from the group consisting of H, F,Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₃)₂, cyclopropyl, OCH₃, OCF₃, CH₂OCH₃ andCH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅ straight chainalkyl; C₂-C₆ branched alkyl, or R₉ and R₁₀ together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7; R₁₃ and R₁₄ are, independently, selected from the group consisting ofH, F, CH₃, C₁-C₆ straight chain alkyl, and C₂-C₆ branched alkyl; and Zis selected from the group consisting of CO₂H, CONHS(O)₂-Aryl,CONHS(O)₂-Alkyl, and

These compounds have one or more of the following characteristics: (i)an inhibition constant (K_(i)) with regard to H1 receptor binding ofless than 500 nM; (ii) a K_(i) with regard to off target binding to anoff target selected from the group consisting of M1, M2, M3, D1, D2, D3,α1 and α2 that is more than 10 times greater than the K_(i) with regardto the H1 receptor; (iii) a nonREM peak time value that is greater than55% nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) and administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.

In one embodiment, the compounds of Formula II have one or more of thefollowing characteristics:

(i) an inhibition constant (K_(i)) with regard to H1 receptor binding ofless than 150 nM; (ii) a K_(i) with regard to off target binding to anoff target selected from the group consisting of M1, M2, and M3, that isgreater than 10 μM; (iii) a nonREM peak time value that is greater than55% nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) acumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 17minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 5 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 6 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) administration of said compound to a subject does notdisproportionately inhibit locomotor activity or motor tone relative tothe normal effects of sleep.

In one embodiment, R₉ and R₁₀ and the carbon they are attached to areabsent. In another embodiment, R₉ and R₁₀, together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7. For example, in one embodiment, R₉ and R₁₀, together with the carbonto which they are attached, are connected to form a spiro cyclopropylring.

In one embodiment, Z is CO₂H or

In one embodiment, o is zero.

In one embodiment, at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at leastone of R₉-R₁₀, are not hydrogen when Z is COOH. In one embodiment, R₁,R₃, R₆-R₇ and R₁₃-R₁₄ are each hydrogen and R₂ is not H, CH₃, CF₃, Cl orBr. In one embodiment, R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ isCH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₁, R₃,R₇, R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂CH₂OH, and R₂ is not H, CH₃,CF₃, Cl or Br. In one embodiment, each of R₁, R₃, R₆, R₇, R₁₃ and R₁₄ isH and R₂ is F. In one embodiment, each of R₁, R₃, R₇, R₁₃ and R₁₄ is H,R₆ is OCH₃, and R₂ is OCH₃. In one embodiment, each of R₁, R₃, R₇, R₁₃and R₁₄ is H, R₆ is F, and R₂ is OCH₃.

In one embodiment, at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen. In one embodiment, at least one of R₁₃-R₁₄ is not hydrogen. Inone embodiment, R₃ is not hydrogen. In one embodiment, R₇ is nothydrogen.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl.

In one embodiment, the sleep modulation is, e.g., decreasing the time tosleep onset, increasing the average sleep bout length, and/or increasingthe maximum sleep bout length. In one embodiment, the sleep modulationtreats a sleep disorder.

Pharmaceutical compositions that include a compound of Formula II orpharmaceutically acceptable salt thereof are also used in the methods ofmodulating sleep in a subject. In one embodiment, the compound ofFormula II or pharmaceutically acceptable salt thereof isco-administered with one or more additional therapies.

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount of acompound having the formula of Formula III:

or a pharmaceutically effective salt thereof, wherein m and n are,individually, 0-4, X is absent, O, S, C(O), SO or SO₂; R₁ is H, F, Cl,Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, cyclopropyl, CH₂OCH₂CH₃, CH₂OCH₃, CH₂OCH₂CH₃, or OCH₃; R₂, R₃, R₆, and R₇ are, independently, selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, OCH₃,CH₂OCH₃, and CH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅straight chain alkyl; C₂-C₆ branched alkyl, or R₉, and R₁₀, togetherwith the carbon to which they are attached, are connected to form aspiro ring of size 3-7; R₁₃ and R₁₄ are, independently, selected fromthe group consisting of H, F, CH₃, C₁-C₆ straight chain alkyl, C₂-C₆branched alkyl, and CH₂OCH₃; and Z is selected from the group consistingof CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, and

These compounds of Formula III have one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 500 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, M3, D1, D2, D3, α1 and α2 that is more than 10 times greaterthan the K_(i) with regard to the H1 receptor; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 13 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 3 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 5 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) and administration of said compound to asubject does not disproportionately inhibit locomotor activity relativeto the normal effects of sleep.

In one embodiment, the compound has one or more of the followingcharacteristics:

(i) an inhibition constant (K_(i)) with regard to H1 receptor binding ofless than 150 nM; (ii) a K_(i) with regard to off target binding to anoff target selected from the group consisting of M1, M2, and M3, that isgreater than 10 μM; (iii) a nonREM peak time value that is greater than55% nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 17minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 5 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 6 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) administration of said compound to a subject does notdisproportionately inhibit locomotor activity or motor tone relative tothe normal effects of sleep.

In one embodiment, R₃ is H. In one embodiment, R₉ and R₁₀, together withthe carbon to which they are attached, are connected to form a spiroring of size 3-7. For example, in one embodiment, R₉ and R₁₀, togetherwith the carbon to which they are attached, are connected to form aspiro cyclopropyl ring.

In one embodiment, Z is CO₂H or

In one embodiment, at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at leastone of R₉-R₁₀, are not hydrogen when Z is COOH.

In one embodiment, R₁, R₃, R₆-R₇ and R₁₃-R₁₄ are each hydrogen, and R₂is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₁, R₃, R₇, R₁₃ and R₁₄are each hydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. Inone embodiment, R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ isCH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, eachof R₁, R₃, R₆, R₇, R₁₃ and R₁₄ is H, and R₂ is F. In one embodiment,each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is OCH₃, and R₂ is OCH₃. In oneembodiment, each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is F, and R₂ isOCH₃.

In one embodiment, at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen. In one embodiment, at least one of R₁₃-R₁₄ is not hydrogen. Inone embodiment, R₃ is not hydrogen. In one embodiment, R₇ is nothydrogen.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl.

In one embodiment, the sleep modulation is, e.g., decreasing the time tosleep onset, increasing the average sleep bout length, and/or increasingthe maximum sleep bout length. In one embodiment, the sleep modulationtreats a sleep disorder.

Pharmaceutical compositions that include a compound of Formula III orpharmaceutically acceptable salt thereof are also used in the methods ofmodulating sleep according to the invention.

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount of acompound having the formula of Formula IV:

or a pharmaceutically effective salt thereof wherein t is 0-6; R₂ and R₆are, independently, H, F, Cl, Br, CF₃, CH₃, OH, OCH₃, CH₂OCH₃, orCH₂OCH₂CH₃; R₉-R₁₀ are H, CH₃ CH₂CH₃, or R₉ and R₁₀, together with thecarbon to which they are attached, are connected to form a spiro ring ofsize 3 to 7; and Z is CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl or

provided that when Z is COOH, t does not equal zero. These compounds ofFormula IV have one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 500 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, M3, D1, D2, D3, α1and α2 that is more than 10 times greater than the K_(i) with regard tothe H1 receptor; (iii) a nonREM peak time value that is greater than 55%nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) and administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.

In one embodiment, the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 150 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, and M3, that is greater than 10 μM; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 17 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 5 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 6 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity or motor tonerelative to the normal effects of sleep.

In one embodiment, the compound of Formula IV is IVa, IVb, IVc, or IVd.In one embodiment, R₉ and R₁₀ together with the carbon to which they areattached, are connected to form a spiro ring of size 3 to 7. Forexample, in one embodiment, R₉ and R₁₀ together with the carbon to whichthey are attached, are connected to form a spiro 3-membered cyclopropylring.

In one embodiment, Z is CO₂H or

In one embodiment, at least one of R₂, and R₆ and at least one ofR₉-R₁₀, are not hydrogen when Z is COOH. In one embodiment, R₆ ishydrogen, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₆ isCH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₆ isCH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₆ isH, and R₂ is F. In one embodiment, R₆ is OCH₃, and R₂ is OCH₃. In oneembodiment, R₆ is F, and R₂ is OCH₃.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl.

In one embodiment, the sleep modulation is selected from the groupconsisting of decreasing the time to sleep onset, increasing the averagesleep bout length, and increasing the maximum sleep bout length. In oneembodiment, the sleep modulation treats a sleep disorder.

Pharmaceutical compositions that include a compound of Formula IV orpharmaceutically acceptable salt thereof are also used in the methods ofmodulating sleep according to the invention.

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 10a:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 47a:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 49a:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 49b:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 37:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 38a:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 38b:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 39a:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 39b:

In another aspect, the invention provides a compound according toFormula I:

or a pharmaceutically effective salt thereof, wherein m n, o, p, q are,individually, 0-6; X and Y are, individually, absent, O, S, C(O), SO orSO₂; R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are, independently selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chainalkyl, C₃-C₆ branched alkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃,OCF₃, CH₂OCH₃, CH₂CH₂OCH₃, CH₂OCH₂CH₃, and C₁-C₆ hydroxyalkyl; anyhydrogen in the CH₂ groups in the linker is optionally substituted withH, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chain alkyl, C₃-C₆ branchedalkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃, OCF₃, CH₂OCH₃,CH₂CH₂OCH₃, CH₂OCH₂CH₃, or C₁-C₆ hydroxyalkyl; R₉, R₁₀, R₁₁, and R₁₂are, independently, H, C₁-C₅ straight chain alkyl, C₂-C₆ branched alkyl,R₉ and R₁₀ together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to 7, or R₁₁ and R₁₂ togetherwith the carbon to which they are attached, are connected to form aspiro ring of size 3 to 7; or substituents on two different atoms areconnected to form a ring of size 3 to 7; R₁₃ and R₁₄ are, independently,selected from the group consisting of H, F, Cl, Br, CH₃, C₁-C₆ straightchain alkyl, and C₂-C₆ branched alkyl; and Z is selected from the groupconsisting of CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl,CONHS(O)₂-Heteroaryl, SO₃H, SO₂H, S(O)₂NHCO-alkyl, S(O)₂NHCO-aryl,S(O)NHCO-alkyl, S(O)NHCO-aryl, P(O)(OH)₂, P(O)OH,

the compound has one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 500 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, M3, D1, D2, D3, α1and α2 that is more than 10 times greater than the K_(i) with regard tothe H1 receptor; (iii) a nonREM peak time value that is greater than 55%nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) and administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.

In one embodiment, the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 150 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, and M3, that is greater than 10 μM; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 17 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 5 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 6 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity or motor tonerelative to the normal effects of sleep.

The methods of the invention are used to treat a variety of subjects,including, for example, humans, companion animals, farm animals,laboratory animals and wild animals.

In one embodiment, the compound used in this method of modulating sleepis 47a, 47b, 47c, 47d, 49a, 49b, 49c, 49d, 10a, 10b, 10c, 10d, 37, 38a,38b, 39a or 39b.

In one embodiment, the R₉ and R₁₀ and the carbon they are attached toare absent. In one embodiment, R₁₁ and R₁₂, together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7. For example, R₁₁ and R₁₂ together with the carbon to which they areattached, are connected to form a spiro 3-membered cyclopropyl ring.

In one embodiment, Z is CO₂H or

In one embodiment where Z is COOH, at least one of R₁-R₈, R₁₃-R₁₄ and atleast one of R₉-R₁₀, R₁₁-R₁₂ are not hydrogen.

In one embodiment, R₁, R₃-R₈ and R₁₃-R₁₄ are each hydrogen and R₂ is notH, CH₃, CF₃, Cl or Br. In another embodiment, R₁, R₃-R₅, R₇-R₈ andR₁₃-R₁₄ are each hydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃, CF₃, Cl orBr. In one embodiment, R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ are each hydrogen,R₆ is CH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In anotherembodiment, each of R₁, R₃-R₈ and R₁₃-R₁₄ is H and R₂ is F. In oneembodiment, each of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ is H, R₆ is OCH₃, andR₂ is OCH₃. In another embodiment, each of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄is H, R₆ is F, and R₂ is OCH₃.

In one embodiment, at least three of R₁-R₈ are not hydrogen. In oneembodiment, at least one of R₁₃-R₁₄ is not hydrogen. In one embodiment,R₃ is not hydrogen. In one embodiment, R₇ is not hydrogen.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl. In one embodiment, R₁₁, and R₁₂ are each ethyl.In another embodiment, R₁₁, and R₁₂ are each ethyl.

In one embodiment, R₁₁ and R₁₂ and the carbon to which they are attachedare connected to form a spiro ring of size 3-7. For example, in oneembodiment, R₁₁ and R₁₂ and the carbon to which they are attached areconnected to form a three-membered spiro (cyclopropyl) ring. In oneaspect, the compound of Formula I also includes a pharmaceuticallyacceptable excipient.

In another aspect, the invention provides a compound of Formula II:

or a pharmaceutically effective salt thereof, wherein m, n, and o are,individually, 0-6; X is absent, O, S, C(O), SO or SO₂; R₁, R₂, R₃, R₆,and R₇ are, independently selected from the group consisting of H, F,Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₃)₂, cyclopropyl, OCH₃, OCF₃, CH₂OCH₃ andCH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅ straight chainalkyl; C₂-C₆ branched alkyl, or R₉ and R₁₀ together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7; R₁₃ and R₁₄ are, independently, selected from the group consisting ofH, F, CH₃, C₁-C₆ straight chain alkyl, and C₂-C₆ branched alkyl; and Zis selected from the group consisting of CO₂H, CONHS(O)₂-Aryl,CONHS(O)₂-Alkyl, and

These compounds have one or more of the following characteristics: (i)an inhibition constant (K_(i)) with regard to H1 receptor binding ofless than 500 nM; (ii) a K_(i) with regard to off target binding to anoff target selected from the group consisting of M1, M2, M3, D1, D2, D3,α1 and α2 that is more than 10 times greater than the K_(i) with regardto the H1 receptor; (iii) a nonREM peak time value that is greater than55% nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) and administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.

In one embodiment, the compounds of Formula II have one or more of thefollowing characteristics: (i) an inhibition constant (K_(i)) withregard to H1 receptor binding of less than 150 nM; (ii) a K_(i) withregard to off target binding to an off target selected from the groupconsisting of M1, M2, and M3, that is greater than 10 μM; (iii) a nonREMpeak time value that is greater than 55% nonREM sleep per hour by thethird hour after said compound is administered to a subject; (iv) acumulative total increase in nonREM sleep not less than 20 minutes forcompound doses that produce maximum sleep consolidation; (v) a longestsleep bout that is greater than 17 minutes in duration; (vi) net longestsleep bout post treatment is greater than or equal to 5 minutes whenadjusted using a baseline value obtained at least 24 hours prior toadministration of said compound to a subject; (vii) an average sleepbout that is greater than 6 minutes at absolute peak; (viii)administration of said compound to a subject does not produceappreciable amounts of rebound insomnia; (ix) administration of saidcompound to a subject does not appreciably inhibit REM sleep; and (x)administration of said compound to a subject does not disproportionatelyinhibit locomotor activity or motor tone relative to the normal effectsof sleep.

In one embodiment, R₉ and R₁₀ and the carbon they are attached to areabsent. In another embodiment, R₉ and R₁₀, together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7. For example, in one embodiment, R₉ and R₁₀, together with the carbonto which they are attached, are connected to form a spiro cyclopropylring.

In one embodiment, Z is CO₂H or

In one embodiment, o is zero.

In one embodiment, at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at leastone of R₉-R₁₀, are not hydrogen when Z is COOH. In one embodiment, R₁,R₃, R₆-R₇ and R₁₃-R₁₄ are each hydrogen and R₂ is not H, CH₃, CF₃, Cl orBr. In one embodiment, R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ isCH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₁, R₃,R₇, R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂CH₂OH, and R₂ is not H, CH₃,CF₃, Cl or Br. In one embodiment, each of R₁, R₃, R₆, R₇, R₁₃ and R₁₄ isH and R₂ is F. In one embodiment, each of R₁, R₃, R₇, R₁₃ and R₁₄ is H,R₆ is OCH₃, and R₂ is OCH₃. In one embodiment, each of R₁, R₃, R₇, R₁₃and R₁₄ is H, R₆ is F, and R₂ is OCH₃.

In one embodiment, at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen. In one embodiment, at least one of R₁₃-R₁₄ is not hydrogen. Inone embodiment, R₃ is not hydrogen. In one embodiment, R₇ is nothydrogen.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl.

In another aspect, the invention provides a compound of Formula III:

or a pharmaceutically effective salt thereof, wherein m and n are,individually, 0-4, X is absent, O, S, C(O), SO or SO₂; R₁ is H, F, Cl,Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, cyclopropyl, CH₂OCH₂CH₃, CH₂OCH₃, CH₂OCH₂CH₃, or OCH₃; R₂, R₃, R₆, and R₇ are, independently, selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, OCH₃,CH₂OCH₃, and CH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅straight chain alkyl; C₂-C₆ branched alkyl, or R₉, and R₁₀, togetherwith the carbon to which they are attached, are connected to form aspiro ring of size 3-7; R₁₃ and R₁₄ are, independently, selected fromthe group consisting of H, F, CH₃, C₁-C₆ straight chain alkyl, C₂-C₆branched alkyl, and CH₂OCH₃; and Z is selected from the group consistingof CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, and

These compounds of Formula III have one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 500 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, M3, D1, D2, D3, α1 and α2 that is more than 10 times greaterthan the K_(i) with regard to the H1 receptor; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 13 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 3 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 5 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) and administration of said compound to asubject does not disproportionately inhibit locomotor activity relativeto the normal effects of sleep.

In one embodiment, the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 150 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, and M3, that is greater than 10 μM; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 17 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 5 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 6 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity or motor tonerelative to the normal effects of sleep.

In one embodiment, R₃ is H. In one embodiment, R₉ and R₁₀, together withthe carbon to which they are attached, are connected to form a spiroring of size 3-7. For example, in one embodiment, R₉ and R₁₀, togetherwith the carbon to which they are attached, are connected to form aspiro cyclopropyl ring.

In one embodiment, Z is CO₂H or

In one embodiment, at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at leastone of R₉-R₁₀, are not hydrogen when Z is COOH.

In one embodiment, R₁, R₃, R₆-R₇ and R₁₃-R₁₄ are each hydrogen, and R₂is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₁, R₃, R₇, R₁₃ and R₁₄are each hydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. Inone embodiment, R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ isCH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, eachof R₁, R₃, R₆, R₇, R₁₃ and R₁₄ is H, and R₂ is F. In one embodiment,each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is OCH₃, and R₂ is OCH₃. In oneembodiment, each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is F, and R₂ isOCH₃.

In one embodiment, at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen. In one embodiment, at least one of R₁₃-R₁₄ is not hydrogen. Inone embodiment, R₃ is not hydrogen. In one embodiment, R₇ is nothydrogen.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl.

In another aspect, the invention provides a compound of Formula IV:

or a pharmaceutically effective salt thereof wherein t is 0-6; R₂ and R₆are, independently, H, F, Cl, Br, CF₃, CH₃, OH, OCH₃, CH₂OCH₃, orCH₂OCH₂CH₃; R₉-R₁₀ are H, CH₃ CH₂CH₃, or R₉ and R₁₀, together with thecarbon to which they are attached, are connected to form a spiro ring ofsize 3 to 7; and Z is CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl or

provided that when Z is COOH, t does not equal zero. The compounds ofFormula IV have one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 500 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, M3, D1, D2, D3, α1and α2 that is more than 10 times greater than the K_(i) with regard tothe H1 receptor; (iii) a nonREM peak time value that is greater than 55%nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) and administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.

In one embodiment, the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 150 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, and M3, that is greater than 10 μM; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 17 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 5 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 6 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity or motor tonerelative to the normal effects of sleep.

In one embodiment, the compound of Formula IV is IVa, IVb, IVc, or IVd.In one embodiment, R₉ and R₁₀ together with the carbon to which they areattached, are connected to form a spiro ring of size 3 to 7. Forexample, in one embodiment, R₉ and R₁₀ together with the carbon to whichthey are attached, are connected to form a spiro 3-membered cyclopropylring.

In one embodiment, Z is CO₂H or

In one embodiment, at least one of R₂, and R₆ and at least one ofR₉-R₁₀, are not hydrogen when Z is COOH. In one embodiment, R₆ ishydrogen, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₆ isCH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₆ isCH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br. In one embodiment, R₆ isH, and R₂ is F. In one embodiment, R₆ is OCH₃, and R₂ is OCH₃. In oneembodiment, R₆ is F, and R₂ is OCH₃.

In one embodiment, R₉ and R₁₀ are each methyl. In another embodiment, R₉and R₁₀ are each ethyl.

In one embodiment, the sleep modulation is selected from the groupconsisting of decreasing the time to sleep onset, increasing the averagesleep bout length, and increasing the maximum sleep bout length. In oneembodiment, the sleep modulation treats a sleep disorder.

Pharmaceutical compositions that include a compound of Formula IV orpharmaceutically acceptable salt thereof are also used in the methods ofmodulating sleep according to the invention.

In another aspect, the invention provides a compound having thestructure of compound 10a:

In another aspect, the invention provides a compound having thestructure of compound 47a:

In another aspect, the invention provides a compound having thestructure of compound 49a:

In another aspect, the invention provides a compound having thestructure of compound 49b:

In another aspect, the invention provides a compound having thestructure of compound 37:

In another aspect, the invention provides a compound having thestructure of compound 38a:

In another aspect, the invention provides a compound having thestructure of compound 38b:

In another aspect, the invention provides a compound having thestructure of compound 39a:

In another aspect, the invention provides a method of modulating sleepin a subject by administering a therapeutically effective amount ofcompound 39b:

In one aspect, the doxepin analogs of the invention are used in thetreatment of a sleep disorder, including, for example, circadian rhythmabnormality, insomnia, parasomnia, sleep apnea syndrome, narcolepsy andhypersomnia.

In one embodiment, the doxepin analogs of the invention are used in thetreatment of a circadian rhythm abnormality, such as, for example, jetlag, shift-work disorders, delayed sleep phase syndrome, advanced sleepphase syndrome and non-24 hour sleep-wake disorder.

In another embodiment, the doxepin analogs are used in the treatment ofinsomnia, including, for example, extrinsic insomnia, psychophysiologicinsomnia, altitude insomnia, restless leg syndrome, periodic limbmovement disorder, medication-dependent insomnia, drug-dependentinsomnia, alcohol-dependent insomnia and insomnia associated with mentaldisorders, such as anxiety. The compounds of the invention are also usedto treat sleep fragmentation associated with Parkinson's disease,Alzheimer's disease, Huntington's disease, and other dystonias.

In one embodiment, the doxepin analogs of the invention are used totreat a parasomnia disorder, such as, e.g., somnambulism, pavornocturnus, REM sleep behavior disorder, sleep bruxism and sleepenuresis.

In another embodiment, the doxepin analogs are used to treat a sleepapnea disorder, such as, for example, central sleep apnea, obstructivesleep apnea and mixed sleep apnea.

In another embodiment, the doxepin analogs are used to treat disordersrelated to sleep disorders, such as, for example, fibromyalgia.

The above description sets forth rather broadly the more importantfeatures of the present invention in order that the detailed descriptionthereof that follows may be understood, and in order that the presentcontributions to the art may be better appreciated. Other objects andfeatures of the present invention will become apparent from thefollowing detailed description considered in conjunction with theexamples.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph depicting typical hERG current tracings recorded at22° C. for a vehicle control and a positive control.

FIG. 2 is a graph depicting the binding curves for compound 37 andtriprolidine binding to the H1 receptor.

FIG. 3 is a graph depicting the binding curves for compound 37 and(−)-scopolamine, MeBr for the M1 receptor.

FIG. 4 is a graph depicting the binding curves for compound 37 and(−)-scopolamine, MeBr for the M2 receptor.

FIG. 5 is a graph depicting the binding curves for compound 37 and(−)-scopolamine, MeBr for the M3 receptor.

FIG. 6 is a graph depicting the sleep consolidating effects of compound37 (HY10073) administered at a concentration of 30 mg/kg at CT-18.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The details of one or more embodiments of the invention are set forth inthe accompanying description below. Although any methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, the preferred methods andmaterials are now described. Other features, objects, and advantages ofthe invention will be apparent from the description. In thespecification, the singular forms also include the plural unless thecontext clearly dictates otherwise. Unless defined otherwise, alltechnical and scientific terms used herein have the same meaning ascommonly understood by one of ordinary skill in the art to which thisinvention belongs. In the case of conflict, the present specificationwill control.

Definitions

For convenience, certain terms used in the specification, examples andappended claims are collected here.

“Treating”, includes any effect, e.g., lessening, reducing, modulating,or eliminating, that results in the improvement of the condition,disease, disorder, etc.

“Alkyl” includes saturated aliphatic groups, including straight-chainalkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl,octyl, nonyl, decyl), branched-chain alkyl groups (e.g., isopropyl,tert-butyl, isobutyl), cycloalkyl (e.g., alicyclic) groups (e.g.,cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkylsubstituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.“Alkyl” further includes alkyl groups that have oxygen, nitrogen, sulfuror phosphorous atoms replacing one or more hydrocarbon backbone carbonatoms. In certain embodiments, a straight chain or branched chain alkylhas six or fewer carbon atoms in its backbone (e.g., C₁-C₆ for straightchain, C₃-C₆ for branched chain), and more preferably four or fewer.Likewise, preferred cycloalkyls have from three to eight carbon atoms intheir ring structure, and more preferably have five or six carbons inthe ring structure. “C₁-C₆” includes alkyl groups containing one to sixcarbon atoms.

The term “alkyl” also includes both “unsubstituted alkyls” and“substituted alkyls”, the latter of which refers to alkyl moietieshaving substituents replacing a hydrogen on one or more carbons of thehydrocarbon backbone. Such substituents can include, for example, alkyl,alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Cycloalkyls can be further substituted, e.g.,with the substituents described above. An “alkylaryl” or an “aralkyl”moiety is an alkyl substituted with an aryl (e.g., phenylmethyl(benzyl)). “Alkyl” also includes the side chains of natural andunnatural amino acids.

“Aryl” includes groups with aromaticity, including 5- and 6-membered“unconjugated”, or single-ring, aromatic groups that may include fromzero to four heteroatoms, as well as “conjugated”, or multicyclic,systems with at least one aromatic ring. Examples of aryl groups includebenzene, phenyl, pyrrole, furan, thiophene, thiazole, isothiazole,imidazole, triazole, tetrazole, pyrazole, oxazole, isooxazole, pyridine,pyrazine, pyridazine, and pyrimidine, and the like. Furthermore, theterm “aryl” includes multicyclic aryl groups, e.g., tricyclic, bicyclic,e.g., naphthalene, benzoxazole, benzodioxazole, benzothiazole,benzoimidazole, benzothiophene, methylenedioxyphenyl, quinoline,isoquinoline, napthridine, indole, benzofuran, purine, benzofuran,deazapurine, or indolizine. Those aryl groups having heteroatoms in thering structure may also be referred to as “aryl heterocycles”,“heterocycles,” “heteroaryls” or “heteroaromatics”. The aromatic ringcan be substituted at one or more ring positions with such substituentsas described above, as for example, halogen, hydroxyl, alkoxy,alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl,aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylthiocarbonyl, phosphate, phosphonato, phosphinato, cyano, amino(including alkylamino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety. Arylgroups can also be fused or bridged with alicyclic or heterocyclicrings, which are not aromatic so as to form a multicyclic system (e.g.,tetralin, methylenedioxyphenyl).

“Alkenyl” includes unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but that contain atleast one double bond. For example, the term “alkenyl” includesstraight-chain alkenyl groups (e.g., ethenyl, propenyl, butenyl,pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched-chainalkenyl groups, cycloalkenyl (e.g., alicyclic) groups (e.g.,cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl,cyclooctenyl), alkyl or alkenyl substituted cycloalkenyl groups, andcycloalkyl or cycloalkenyl substituted alkenyl groups. The term“alkenyl” further includes alkenyl groups, which include oxygen,nitrogen, sulfur or phosphorous atoms replacing one or more hydrocarbonbackbone carbons. In certain embodiments, a straight chain or branchedchain alkenyl group has six or fewer carbon atoms in its backbone (e.g.,C₂-C₆ for straight chain, C₃-C₆ for branched chain.) Likewise,cycloalkenyl groups may have from three to eight carbon atoms in theirring structure, and more preferably have five or six carbons in the ringstructure. The term “C₂-C₆” includes alkenyl groups containing two tosix carbon atoms.

The term “alkenyl” also includes both “unsubstituted alkenyls” and“substituted alkenyls”, the latter of which refers to alkenyl moietieshaving substituents replacing a hydrogen on one or more hydrocarbonbackbone carbon atoms. Such substituents can include, for example, alkylgroups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

“Alkynyl” includes unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but which containat least one triple bond. For example, “alkynyl” includes straight-chainalkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl,heptynyl, octynyl, nonynyl, decynyl), branched-chain alkynyl groups, andcycloalkyl or cycloalkenyl substituted alkynyl groups. The term“alkynyl” further includes alkynyl groups having oxygen, nitrogen,sulfur or phosphorous atoms replacing one or more hydrocarbon backbonecarbons. In certain embodiments, a straight chain or branched chainalkynyl group has six or fewer carbon atoms in its backbone (e.g., C₂-C₆for straight chain, C₃-C₆ for branched chain). The term “C₂-C₆” includesalkynyl groups containing two to six carbon atoms.

The term “alkynyl” also includes both “unsubstituted alkynyls” and“substituted alkynyls”, the latter of which refers to alkynyl moietieshaving substituents replacing a hydrogen on one or more hydrocarbonbackbone carbon atoms. Such substituents can include, for example, alkylgroups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

Unless the number of carbons is otherwise specified, “lower alkyl”includes an alkyl group, as defined above, but having from one to ten,more preferably from one to six, carbon atoms in its backbone structure.“Lower alkenyl” and “lower alkynyl” have chain lengths of, for example,2-5 carbon atoms.

“Acyl” includes compounds and moieties that contain the acyl radical(CH₃CO—) or a carbonyl group. “Substituted acyl” includes acyl groupswhere one or more of the hydrogen atoms are replaced by for example,alkyl groups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

“Acylamino” includes moieties wherein an acyl moiety is bonded to anamino group. For example, the term includes alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido groups.

“Aroyl” includes compounds and moieties with an aryl or heteroaromaticmoiety bound to a carbonyl group. Examples of aroyl groups includephenylcarboxy, naphthyl carboxy, etc. “Alkoxyalkyl”, “alkylaminoalkyl”and “tioalkoxyalkyl” include alkyl groups, as described above, whichfurther include oxygen, nitrogen or sulfur atoms replacing one or morehydrocarbon backbone carbon atoms, e.g., oxygen, nitrogen or sulfuratoms.

The term “alkoxy” or “alkoxyl” includes substituted and unsubstitutedalkyl, alkenyl, and alkynyl groups covalently linked to an oxygen atom.Examples of alkoxy groups (or alkoxyl radicals) include methoxy, ethoxy,isopropyloxy, propoxy, butoxy, and pentoxy groups. Examples ofsubstituted alkoxy groups include halogenated alkoxy groups. The alkoxygroups can be substituted with groups such as alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkylamino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulflhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.Examples of halogen substituted alkoxy groups include, but are notlimited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,chloromethoxy, dichloromethoxy, and trichloromethoxy.

The terms “heterocyclyl” or “heterocyclic group” include closed ringstructures, e.g., 3- to 10-, or 4- to 7-membered rings, which includeone or more heteroatoms. “Heteroatom” includes atoms of any elementother than carbon or hydrogen. Examples of heteroatoms include nitrogen,oxygen, sulfur and phosphorus.

Heterocyclyl groups can be saturated or unsaturated and includepyrrolidine, oxolane, thiolane, piperidine, piperazine, morpholine,lactones, lactams such as azetidinones and pyrrolidinones, sultams, andsultones. Heterocyclic groups such as pyrrole and furan can havearomatic character. They include fused ring structures such as quinolineand isoquinoline. Other examples of heterocyclic groups include pyridineand purine. The heterocyclic ring can be substituted at one or morepositions with such substituents as described above, as for example,halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,phosphinato, cyano, amino (including alkyl amino, dialkylamino,arylamino, diarylamino, and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, or an aromatic or heteroaromatic moiety. Heterocyclicgroups can also be substituted at one or more constituent atoms with,for example, a lower alkyl, a lower alkenyl, a lower alkoxy, a loweralkylthio, a lower alkylamino, a lower alkylcarboxyl, a nitro, ahydroxyl, —CF₃, or —CN, or the like.

The term “thiocarbonyl” or “thiocarboxy” includes compounds and moietieswhich contain a carbon connected with a double bond to a sulfur atom.

The term “ether” includes compounds or moieties which contain an oxygenbonded to two different carbon atoms or heteroatoms. For example, theterm includes “alkoxyalkyl” which refers to an alkyl, alkenyl, oralkynyl group covalently bonded to an oxygen atom which is covalentlybonded to another alkyl group.

The term “ester” includes compounds and moieties which contain a carbonor a heteroatom bound to an oxygen atom which is bonded to the carbon ofa carbonyl group. The term “ester” includes alkoxycarboxy groups such asmethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl,pentoxycarbonyl, etc. The alkyl, alkenyl, or alkynyl groups are asdefined above.

The term “thioether” includes compounds and moieties which contain asulfur atom bonded to two different carbon or heteroatoms. Examples ofthioethers include, but are not limited to alkthioalkyls,alkthioalkenyls, and alkthioalkynyls. The term “alkthioalkyls” includecompounds with an alkyl, alkenyl, or alkynyl group bonded to a sulfuratom which is bonded to an alkyl group. Similarly, the term“alkthioalkenyls” and alkthioalkynyls” refer to compounds or moietieswherein an alkyl, alkenyl, or alkynyl group is bonded to a sulfur atomwhich is covalently bonded to an alkynyl group.

The term “hydroxy” or “hydroxyl” includes groups with an —OH or —O—.

The term “halogen” includes fluorine, bromine, chlorine, iodine, etc.The term “perhalogenated” generally refers to a moiety wherein allhydrogens are replaced by halogen atoms.

“Polycyclyl” or “polycyclic radical” refers to two or more cyclic rings(e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/orheterocyclyls) in which two or more carbons are common to two adjoiningrings. Rings that are joined through non-adjacent atoms are termed“bridged” rings. Each of the rings of the polycycle can be substitutedwith such substituents as described above, as for example, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,alkylaminocarbonyl, aralkylaminocarbonyl, alkenylaminocarbonyl,alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl,aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,phosphinato, cyano, amino (including alkylamino, dialkylamino,arylamino, diarylamino, and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkyl, alkylaryl, or anaromatic or heteroaromatic moiety.

An “anionic group,” as used herein, refers to a group that is negativelycharged at physiological pH. Preferred anionic groups includecarboxylate, sulfate, sulfonate, sulfinate, sulfamate, tetrazolyl,phosphate, phosphonate, phosphinate, or phosphorothioate or functionalequivalents thereof. “Functional equivalents” of anionic groups areintended to include bioisosteres, e.g., bioisosteres of a carboxylategroup. Bioisosteres encompass both classical bioisosteric equivalentsand non-classical bioisosteric equivalents. Classical and non-classicalbioisosteres are known in the art (see, e.g., Silverman, R. B. TheOrganic Chemistry of Drug Design and Drug Action, Academic Press, Inc.:San Diego, Calif., 1992, pp. 19-23). A particularly preferred anionicgroup is a carboxylate.

The terms “crystal polymorphs” or “polymorphs” refer to the existence ofmore than one crystal form for a compound, salt or solvate thereof.Crystal polymorphs of the doxepin-analog compounds are prepared bycrystallization under different conditions.

It will be noted that the structure of some of the compounds of theinvention includes asymmetric carbon atoms. It is to be understoodaccordingly that the isomers arising from such asymmetry (e.g., allenantiomers and diastereomers) are included within the scope of theinvention, unless indicated otherwise. Such isomers can be obtained insubstantially pure form by classical separation techniques and bystereochemically controlled synthesis. Furthermore, the structures andother compounds and moieties discussed in this application also includeall tautomers thereof. Alkenes can include either the E- or Z-geometry,where appropriate.

The language “doxepin-like compounds” or “doxepin-analog compounds”“doxepin-like compounds” or “doxepin derivative compounds” is intendedto include analogs of doxepin or antihistamines that include two arylgroups linked to the same atom that are linked through a tricyclic ringsystem, e.g. a seven membered ring (i.e., similar to that of doxepin)linked to a double bond to position 4 of a piperidine ring.

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analog is acompound that is similar or comparable in function and appearance, butnot in structure or origin to the reference compound. For example, thereference compound can be a reference antihistamine such as doxepin, andan analog is a substance possessing a chemical structure or chemicalproperties similar to those of the reference antihistamine.

As defined herein, the term “derivative”, e.g., in the term “doxepinderivatives”, refers to compounds that have a common core structure, andare substituted with various groups as described herein. For example,all of the compounds represented by formulae I-VIc are doxepinderivatives, and have one of formulae I-VIc as a common core.

The term “antihistamine” refers to a compound that binds to a H1receptor and blocks the action of histamine.

As used herein, the term “sleep disorder” includes conditions recognizedby one skilled in the art as sleep disorders, for example, conditionsknown in the art or conditions that are proposed to be sleep disordersor discovered to be sleep disorders. A sleep disorder also arises in asubject that has other medical disorders, diseases, or injuries, or in asubject being treated with other medications or medical treatments,where the subject, as a result, has difficulty falling asleep and/orremaining asleep, or experiences unrefreshing sleep, e.g., the subjectexperiences sleep deprivation.

The term “treating a sleep disorder” also includes treating a sleepdisorder component of other disorders, such as CNS disorders (e.g.,mental or neurological disorders such as anxiety). Additionally, theterm “treating a sleep disorder” includes the beneficial effect ofameliorating other symptoms associated with the disorder.

The term “nonREM peak sleep time” is defined as an absolute peak amountof nonREM sleep per hour post treatment, with drug administrationoccurring at Circadian Time (CT) 18, which is 6 hours after lights offin a nocturnal laboratory rat when housed in a LD 12:12 (12-hours lightand 12 hours dark) light-dark cycle. The nominal criteria of 55% nonREMsleep per hour is equivalent to 33 minutes of nonREM sleep per hour.

As used herein, the term “cumulative nonREM sleep” is defined as the nettotal aggregate increase in the number of minutes of nonREM sleep,measured through out the entire duration of a drug's soporific effect,which typically, but not always occurs in the first 6 hourspost-treatment, adjusted for the net total aggregate number of minutesof nonREM sleep that occurred during the corresponding non-treatmentbaseline times of day recorded 24 hours earlier, relative to likevehicle control treatment.

As defined herein, the term “sleep bout” refers to a discrete episode ofcontinuous or near continuous sleep, comprised of nonREM sleep, REMsleep, or both nonREM and REM sleep stages, delimited prior and afterthe episode by greater than two contiguous 10 second epochs ofwakefulness.

As used herein, the term “longest sleep bout length” is defined as thetotal number of minutes an animal remains asleep (nonREM and/or REMsleep stages) during the single longest sleep episode or “bout” thatoccurred beginning in a given hour post-treatment. The “sleep boutlength” measurement criteria assumes sleep is measured continuously in10 second epochs, and is scored based upon the predominant state,computed or otherwise determined as a discrete sleep stage (where sleepstages are defined as nonREM sleep, REM sleep, or wakefulness) duringthe 10 second interval that defines the epoch.

The term “average sleep bout length” is defined as the average duration(in minutes) of every sleep bout that began in a given hour, independentof the individual duration of each episode or bout.

“Rebound insomnia” is defined as period of rebound, paradoxical, orcompensatory wakefulness that occurs after the sleep promoting effectsof a hypnotic or soporific agent.

“REM sleep inhibition” is defined as the reduction of REM sleep timepost-treatment at CT-18 (6 hours after lights-off; LD 12:12) or at CT-5(5 hours after lights-on; LD 12:12). Compounds that reduce REM sleeptime by greater than 15 minutes (relative to baseline and adjusted forvehicle treatment) when administered at either CT-18 or CT-5 areconsidered unacceptable.

Compared with NREM sleep or wakefulness, REM sleep causes ventilatorydepression and episodic cardiovascular changes. During rebound insomnia,the physiological effects of REM sleep are magnified and interrupt thenormal sleep cycles.

As defined herein, “disproportionate locomotor activity inhibition” is areduction of locomotor activity that exceeds the normal and expectedreduction in behavioral activity attributable to sleep.

“Combination therapy” (or “co-therapy”) includes the administration of acompound of the invention and at least a second agent as part of aspecific treatment regimen intended to provide the beneficial effectfrom the co-action of these therapeutic agents. The beneficial effect ofthe combination includes, but is not limited to, pharmacokinetic orpharmacodynamic co-action resulting from the combination of therapeuticagents. Administration of these therapeutic agents in combinationtypically is carried out over a defined time period (usually minutes,hours, days or weeks depending upon the combination selected).“Combination therapy” may, but generally is not, intended to encompassthe administration of two or more of these therapeutic agents as part ofseparate monotherapy regimens that incidentally and arbitrarily resultin the combinations of the present invention. “Combination therapy” isintended to embrace administration of these therapeutic agents in asequential manner, that is, wherein each therapeutic agent isadministered at a different time, as well as administration of thesetherapeutic agents, or at least two of the therapeutic agents, in asubstantially simultaneous manner. Substantially simultaneousadministration can be accomplished, for example, by administering to thesubject a single capsule having a fixed ratio of each therapeutic agentor in multiple, single capsules for each of the therapeutic agents.Sequential or substantially simultaneous administration of eachtherapeutic agent can be effected by any appropriate route including,but not limited to, oral routes, intravenous routes, intramuscularroutes, and direct absorption through mucous membrane tissues. Thetherapeutic agents can be administered by the same route or by differentroutes. For example, a first therapeutic agent of the combinationselected may be administered by intravenous injection while the othertherapeutic agents of the combination may be administered orally.Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection. The sequence in which the therapeutic agents are administeredis not narrowly critical. “Combination therapy” also embraces theadministration of the therapeutic agents as described above in furthercombination with other biologically active ingredients and non-drugtherapies (e.g., surgery or radiation treatment). Where the combinationtherapy further comprises a non-drug treatment, the non-drug treatmentmay be conducted at any suitable time so long as a beneficial effectfrom the co-action of the combination of the therapeutic agents andnon-drug treatment is achieved. For example, in appropriate cases, thebeneficial effect is still achieved when the non-drug treatment istemporally removed from the administration of the therapeutic agents,perhaps by days or even weeks.

The terms “parenteral administration” and “administered parenterally” asused herein refer to modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intra-arterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The term “pulmonary” as used herein refers to any part, tissue or organwhose primary function is gas exchange with the external environment,e.g., O₂/CO₂ exchange, within a patient. “Pulmonary” typically refers tothe tissues of the respiratory tract. Thus, the phrase “pulmonaryadministration” refers to administering the formulations describedherein to any part, tissue or organ whose primary function is gasexchange with the external environment (e.g., mouth, nose, pharynx,oropharynx, laryngopharynx, larynx, trachea, carina, bronchi,bronchioles, alveoli). For purposes of the present invention,“pulmonary” also includes a tissue or cavity that is contingent to therespiratory tract, in particular, the sinuses.

An “effective amount” of a compound of the disclosed invention is thequantity which, when administered to a subject in need of treatment,ameliorates symptoms arising from a sleep disorder, e.g., results in thesubject falling asleep more rapidly, results in more refreshing sleep,reduces duration or frequency of waking during a sleep period, orreduces the duration, frequency, or intensity of other dyssomnias,parasomnias. The amount of the disclosed compound to be administered toa subject will depend on the particular disorder, the mode ofadministration, co-administered compounds, if any, and thecharacteristics of the subject, such as general health, other diseases,age, sex, genotype, body weight and tolerance to drugs. The skilledartisan will be able to determine appropriate dosages depending on theseand other factors.

A “pharmaceutically acceptable salt” or “salt” of the disclosed compoundis a product of the disclosed compound that contains an ionic bond, andis typically produced by reacting the disclosed compound with either anacid or a base, suitable for administering to a subject.

A “pharmaceutical composition” is a formulation containing the disclosedcompounds in a form suitable for administration to a subject. In apreferred embodiment, the pharmaceutical composition is in bulk or inunit dosage form. The unit dosage form is any of a variety of forms,including, for example, a capsule, an IV bag, a tablet, a single pump onan aerosol inhaler, or a vial. The quantity of active ingredient (e.g.,a formulation of the disclosed compound or salts thereof) in a unit doseof composition is an effective amount and is varied according to theparticular treatment involved. One skilled in the art will appreciatethat it is sometimes necessary to make routine variations to the dosagedepending on the age and condition of the patient. The dosage will alsodepend on the route of administration. A variety of routes arecontemplated, including oral, pulmonary, rectal, parenteral,transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal,intranasal, and the like. Dosage forms for the topical or transdermaladministration of a compound of this invention include powders, sprays,ointments, pastes, creams, lotions, gels, solutions, patches andinhalants. In a preferred embodiment, the active compound is mixed understerile conditions with a pharmaceutically acceptable carrier, and withany preservatives, buffers, or propellants that are required.

The term “flash dose” refers to compound formulations that are rapidlydispersing dosage forms.

The term “immediate release” is defined as a release of compound from adosage form in a relatively brief period of time, generally up to about60 minutes. The term “modified release” is defined to include delayedrelease, extended release, and pulsed release. The term “pulsed release”is defined as a series of releases of drug from a dosage form. The term“sustained release” or “extended release” is defined as continuousrelease of a compound from a dosage form over a prolonged period.

A “subject” includes mammals, e.g., humans, companion animals (e.g.,dogs, cats, birds, and the like), farm animals (e.g., cows, sheep, pigs,horses, fowl, and the like) and laboratory animals (e.g., rats, mice,guinea pigs, birds, and the like). Most preferably, the subject ishuman.

The invention provides a method of modulating sleep by administering aneffective amount of a doxepin analog of the invention, which is a moietythat antagonizes a histamine receptor or a collection of histaminereceptors.

Effective sleep modulators have certain characteristics that correspondwith increased efficacy and decreased side effects. Thesecharacteristics include a desired half-life in a subject, controlledonset of desired sedative effects, and minimal to no detectable effecton psychomotor or other central nervous system (CNS) side effects (e.g.,memory deficits, decreased muscle tone, drooping eyelids or drowsiness).For example, effective sleep modulators have a half life in humans ofless than 7 hours, less than 6 hours, less than 5 hours, less than 4hours, approximately 3 hours, or in the range of 3 to 7 hours.

One approach to developing an effective sleep modulator is strategicallyderivitizing a known compound or family of compounds with sleepmodulating activity. Derivitizing may enhance one or more biologicalproperties to allow a compound to perform in an improved manner.Examples of favorable biological properties include, but are notlimited, to induction of a discrete sleep or hypnotic state, activity ofthe therapeutic compound for a discrete period of time, penetrationthrough the blood brain barrier into the CNS, e.g., resulting fromlipophilicity of substituents or conformational lipophilicity (i.e.,lipophilicity as a result of a particular conformation, such as internalsalt formation between a carboxylate anion and a protonated amine),modulation of the half-life of the therapeutic compound, an alterationof charge, an alteration of pharmacokinetics, an alteration of log P bya value of one or more, increased receptor selectivity, reducedperipheral half-life, the ability to increase dosage, increasedperipheral elimination, decreased anti-muscarinic activity, decreasedanti-cholinergic, and any combination thereof.

Derivitizing results in a variety of effects and alter differentmechanisms of action. For example, in some circumstances, a compoundcontaining a particular functional group, such as, e.g., an ester,carboxylic acid, or alcohol group, possesses an improved selectivity fora desired receptor versus undesired receptors when compared with acompound without this group. In other circumstances, the compoundcontaining the particular functional group is more active as atherapeutic agent for treating sleep disorders than the correspondingcompound without this group. The effect of the derivitized compounddepends on the identity of the addition.

By derivitizing a compound in order to enhance favorable biologicalproperties and decrease undesirable side effects, it is possible toimplement a strategy based on potential mechanistic effects orinteractions. For example, in some compounds, the presence of acarboxylic acid results in the ability to form an intramolecular ionicbond that includes the corresponding carboxylate ion, e.g., zwitterionspecies formation with a nitrogen atom within the compound or saltbridge formation. These interactions result in favorable biologicaleffects such as conformational lipophilicity, i.e., increasedlipophilicity as a result of a particular conformation, such as internalsalt formation between a carboxylate anion and a protonated amine. Suchconformational lipophilicity allows penetration through the blood brainbarrier into the CNS, despite that the presence of two polar ions isgenerally thought to inhibit crossing of the non-polar blood-brainbarrier. Another benefit of the presence of the carboxylic acid is animproved ability of the compound to bind selectively to the desiredreceptor.

Compounds of the invention can also be derivitized to produce prodrugs.“Prodrug” includes a precursor form of the drug which is metabolicallyconverted in vivo to produce the active drug. The invention furthercontemplates the use of prodrugs which are converted in vivo to thesleep modulating compounds used in the methods of the invention (see,e.g., R. B. Silverman, 1992, “The Organic Chemistry of Drug Design andDrug Action”, Academic Press, Chp. 8). Such prodrugs can be used toalter the biodistribution (e.g., to allow compounds which would nottypically cross the blood-brain barrier to cross the blood-brainbarrier) or the pharmacokinetics of the sleep modulating compound. Forexample, an anionic group, e.g., a carboxylate, sulfate or sulfonate,can be esterified, e.g., with an alkyl group (e.g., a methyl group) or aphenyl group, to yield an ester. When the ester is administered to asubject, the ester is cleaved, enzymatically or non-enzymatically,reductively or hydrolytically, to reveal the anionic group. Such anester can be cyclic, e.g., a cyclic sulfate or sulfone, or two or moreanionic moieties may be esterified through a linking group. An anionicgroup can be esterified with moieties (e.g., acyloxymethyl esters) whichare cleaved to reveal an intermediate sleep modulating compound whichsubsequently decomposes to yield the active sleep modulating compound.In one embodiment, the prodrug is a reduced form of a carboxylate,sulfate or sulfonate, e.g., an alcohol or thiol, which is oxidized invivo to the sleep modulating compound. Furthermore, an anionic moietycan be esterified to a group which is actively transported in vivo, orwhich is selectively taken up by target organs.

This strategy is applied to sleep modulating compounds to improve theireffectiveness and safety in clinical use. One group of compounds usefulin modulating sleep is related to doxepin, which is a psychotherapeuticagent belonging to the family of compounds commonly known as tricyclicanti-depressants (“TCAs”). More specifically, doxepin is a mixture ofgeometric isomers of a dibenzoxepin tricyclic compound with thefollowing chemical structures:

Doxepin is recommended for treating psychoneuroses, depression, anxiety,alcoholism, other organic diseases and psychotic depressive disorders.

The present invention relates to doxepin-like compounds and their use tomodulate sleep. In general, in one aspect, the doxepin-like compoundsinclude compounds of Formula I:

-   -   where m n, o, p, q are, individually, 0-6, the CH₂ groups are        optionally branched, and any member of the alkylene linker        (e.g., the portion of the molecule connecting the piperidine        ring with the Z group) is substituted with one or more        substituents; X and Y are, individually, absent or O, S, C(O),        SO, or SO₂; R₁-R₈ are, independently, H, F, Cl, Br, CF₃, CH₃, or        C₂-C₆ straight chain alkyl, C₃-C₆ branched alkyl, C₃-C₇        heteroalkyl, C₃-C₇ cycloalkyl, C₃-C₇ cycloheteroalkyl, each of        which is optionally heteroatom substituted, OCH₃, OCF₃, CH₂OCH₃,        CH₂CH₂OCH₃, CH₂OCH₂CH₃, or C₁-C₆ hydroxyalkyl (branched or        unbranched) and/or cyclic; R₉-R₁₂ are H, C₁-C₅ straight chain or        branched alkyl (optionally containing a heteroatom). Optionally,        substituents on nearby atoms are connected to form a ring of        size 3-7 or substituents on the same atom (i.e., geminal        substituents) are connected to form a ring of size 3-7, e.g., R₉        and R₁₀ (or R₁₁ and R₁₂) are directly bonded to form a spiro        ring of size 3-7; R₁₃ and R₁₄ are independently H, F, Cl, Br,        CH₃, lower alkyl or cyclic lower alkyl, or R₁₃ and R₁₄ are        directly bonded to form a spiro ring of size 3-7; Z is CO₂H,        CONHS(O)₂-Aryl (optionally substituted), CONHS(O)₂-Alkyl        (optionally substituted), CONHS(O)₂-Heteroaryl (optionally        substituted), SO₃H, SO₂H, S(O)₂NHCO-alkyl, S(O)₂NHCO-aryl,        S(O)NHCO-alkyl, S(O)NHCO-aryl, P(O)(OH)₂, P(O)OH,

Where at least one of R₁-R₈, R₁₃-R₁₄ and at least one of R₉-R₁₀, R₁₁-R₁₂are not hydrogen when Z is COOH. Also, when R₁, R₃-R₈ and R₁₃-R₁₄ areeach hydrogen, R₂ is not H, CH₃, CF₃, Cl or Br. When R₁, R₃-R₅, R₇-R₈and R₁₃-R₁₄ are each hydrogen, and R₆ is CH₂OH, R₂ is not H, CH₃, CF₃,Cl or Br. When R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ are each hydrogen, and R₆ isCH₂CH₂OH, R₂ is not H, CH₃, CF₃, Cl or Br.

When each of R₁, R₃-R₈, and R₁₃-R₁₄ is H, R₂ is preferably F. When eachof R₁, R₃-R₅, R₇-R₅ and R₁₃-R₁₄ is H, and R₆ is OCH₃, R₂ is preferablyOCH₃. When each of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ is H, and R₆ is F, R₂ ispreferably OCH₃.

In some embodiments, at least three of R₁-R₅ are not hydrogen. In someembodiments, at least one of R₁₃-R₁₄ is not hydrogen. In one embodiment,R₃ is not hydrogen. In another embodiment, R₇ is not hydrogen.

In some embodiments, R₉ and R₁₀ (along with the carbon they are attachedto) are absent.

In some embodiments, R₉ and R₁₀ are each methyl. In some embodiments, R₉and R₁₀ are each ethyl. In some embodiments, R₁₁ and R₁₂ are eachmethyl. In some embodiments, R₁₁ and R₁₂ are each ethyl. In otherembodiments, R₉ and R₁₀ (along with the carbon they are attached to) areconnected to form a spiro ring of size 3-7. In particular embodiments,R₉ and R₁₀ (along with the carbon they are attached to) are connected toform a three-membered spiro (cyclopropyl) ring. In other embodiments,R₁₁ and R₁₂ (along with the carbon they are attached to) are connectedto form a Spiro ring of size 3-7. In particular embodiments, R₁₁ and R₁₂(along with the carbon they are attached to) are connected to form athree-membered spiro (cyclopropyl) ring.

In general, in another aspect, the present invention relates todoxepin-like compounds having the formula of Formula II:

-   -   where m n, and o, are, individually, 0-6, and the CH₂ groups in        the linker are optionally branched; X is absent or O, S, C(O),        SO, or SO₂; R₁, R₂, R₃, R₆ and R₇ are H, F, Cl, Br, CF₃, CH₃,        CH₂CH₃, CH(CH₃)₂, cyclopropyl, OCH₃, OCF₃, CH₂OCH₃, or        CH₂OCH₂CH₃; R₉-R₁₀ are H, C₁-C₅ straight chain or branched alkyl        (optionally containing a heteroatom), and/or, together with the        atom to which they are attached, are connected to form a ring of        size 3-7; R₁₃ and R₁₄ are independently, H, F, CH₃, lower alkyl        or cyclic lower alkyl, or R₁₃ and R₁₄ are directly bonded to        form a spiro ring of size 3-7. Optionally, substituents on        nearby atoms of the linker are connected to form a ring of size        3-7 or substituents on the same atom (i.e., geminal        substituents) are connected to form a ring of size 3-7, e.g.,        are directly bonded to form a spiro ring of size 3-7. Z is CO₂H,        CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, or

Where at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at least one of R₉-R₁₀,are not hydrogen when Z is COOH. Also, when R₁, R₃, R₆-R₇ and R₁₃-R₁₄are each hydrogen, R₂ is not H, CH₃, CF₃, Cl or Br. When R₁, R₃, R₇, R₁₃and R₁₄ are each hydrogen, and R₆ is CH₂OH, R₂ is not H, CH₃, CF₃, Cl orBr. When R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, and R₆ is CH₂CH₂OH,R₂ is not H, CH₃, CF₃, Cl or Br.

When each of R₁, R₃, R₆, R₇, R₁₃ and R₁₄ is H, R₂ is preferably F. Wheneach of R₁, R₃, R₇, R₁₃ and R₁₄ is H, and R₆ is OCH₃, R₂ is preferablyOCH₃. When each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, and R₆ is F, R₂ ispreferably OCH₃.

In some embodiments, at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen. In some embodiments, at least one of R₁₃-R₁₄ is not hydrogen.In one embodiment, R₃ is not hydrogen. In another embodiment, R₇ is nothydrogen.

In some embodiments, R₉ and R₁₀ are each methyl. In some embodiments, R₉and R₁₀ are each ethyl. In other embodiments, R₉ and R₁₀ (along with thecarbon they are attached to) are connected to form a spiro ring of size3-7. In particular embodiments, R₉ and R₁₀ (along with the carbon theyare attached to) are connected to form a three-membered spiro(cyclopropyl) ring.

In general, in another aspect, the present invention relates todoxepin-like compounds having the formula of Formula III:

-   -   where m and n are, individually, 0-4, and the CH₂ moieties are        optionally branched; X is absent or O, S, C(O), SO, or SO₂;        R₉-R₁₀ are H, C₁-C₃ alkyl, optionally with heteroatom        substitution, branching and/or connected to form a ring of size        3-5; R₁₃ and R₁₄ are independently H, F, CH₃, lower alkyl or        cyclic lower alkyl, CH₂OCH₃, or R₁₃ and R₁₄ are directly bonded        to form a spiro ring of size three to six; R₁ is H, F, Cl, Br,        CF₃, CH₃, CH₂CH₃, CH(CH₂)₃, cyclopropyl, OCH₃, CH₂OCH₃ or        CH₂OCH₂CH₃; R₂, R₃, R₆, and R₇ are, independently, H, F, Br,        CF₃, CH₃, OCH₃, CH₂OCH₃ or CH₂OCH₂CH₃; Z is CO₂H,        CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, or        Optionally, substituents on nearby atoms of the linker are        connected to form a ring of size 3-7.

Where at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at least one of R₉-R₁₀,are not hydrogen when Z is COOH. Also, when R₁, R₃, R₆-R₇ and R₁₃-R₁₄are each hydrogen, R₂ is not H, CH₃, CF₃, Cl or Br. When R₁, R₃, R₇, R₁₃and R₁₄ are each hydrogen, and R₆ is CH₂OH, R₂ is not H, CH₃, CF₃, Cl orBr. When R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, and R₆ is CH₂CH₂OH,R₂ is not H, CH₃, CF₃, Cl or Br.

When each of R₁, R₃, R₆, R₇, R₁₃ and R₁₄ is H, R₂ is preferably F. Wheneach of R₁, R₃, R₇, R₁₃ and R₁₄ is H, and R₆ is OCH₃, R₂ is preferablyOCH₃. When each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, and R₆ is F, R₂ ispreferably OCH₃.

In some embodiments, at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen. In some embodiments, at least one of R₁₃-R₁₄ is not hydrogen.In one embodiment, R₃ is not hydrogen. In another embodiment, R₇ is nothydrogen.

In some embodiments, R₉ and R₁₀ are each methyl. In some embodiments, R₉and R₁₀ are each ethyl. In other embodiments, R₉ and R₁₀ (along with thecarbon they are attached to) are connected to form a spiro ring of size3-7. In particular embodiments, R₉ and R₁₀ (along with the carbon theyare attached to) are connected to form a three-membered spiro(cyclopropyl) ring.

In another aspect, the present invention relates to doxepin-likecompounds having the formula of Formula IV:

-   -   where t is between 0 and 6; R₂ and R₆ are, independently, H, F,        Cl, Br, CF₃, CH₃, OH, OCH₃, CH₂OCH₃, CH₂OCH₂CH₃; R₉-R₁₀ are H,        CH₃ or CH₂CH₃, and are optionally connected to form a spiro ring        of size 3 to 7; and Z is CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl        or        provided that when Z is COOH, t is not zero.

At least one of R₂, and R₆ and at least one of R₉-R₁₀, are not hydrogenwhen Z is COOH. Also, when R₆ is hydrogen, R₂ is not H, CH₃, CF₃, Cl orBr. When R₆ is CH₂OH, R₂ is not H, CH₃, CF₃, Cl or Br. When R₆ isCH₂CH₂OH, R₂ is not H, CH₃, CF₃, Cl or Br.

When R₆ is H, R₂ is preferably F. When R₆ is OCH₃, R₂ is preferablyOCH₃. When R₆ is F, R₂ is preferably OCH₃.

In some embodiments, R₉ and R₁₀ are each methyl. In some embodiments, R₉and R₁₀ are each ethyl. In other embodiments, R₉ and R₁₀ (along with thecarbon they are attached to) are connected to form a spiro ring of size3-7. In particular embodiments, R₉ and R₁₀ (along with the carbon theyare attached to) are connected to form a three-membered spiro(cyclopropyl) ring.

For example, when R₉ and R₁₀ are methyl, compounds have the generalformula IVa:

-   -   when R₉ and R₁₀ are connected to form a 3 membered spiro ring        (cyclopropyl), compounds have the general formula IVb:    -   when R₉ and R₁₀ are ethyl, compounds have the general formula        IVc:    -   when R₉ and R₁₀ are ethyl, and the Cl carbons are connected to        form a 3 membered Spiro ring (cyclopropyl), compounds have the        general formula IVd:    -   and when and R₉ and R₁₀ are hydrogen, compounds have the general        formula IVe:

Some representative compounds of the invention are shown in Table 1.TABLE 1 Di-, mono- and un- substituted doxepin derivatives Compound R₂R₆ R₉, R₁₀ t size ring Z 46a F CH₃O CH₃ 1 none COOH 47a CH₃O F CH₃ 1none COOH 47c CH₃O F CH₂CH₃ 1 none COOH 49b F H CH₃ 1 3 COOH 49a F H CH₃1 none COOH 49c F H CH₂CH₃ 1 none COOH 50b CH₃O H CH₃ 1 3 COOH 50a CH₃OH CH₃ 1 none COOH 51b Cl H CH₃ 1 3 COOH 52a H CH₃O CH₃ 1 none COOH 52b HCH₃O CH₃ 1 3 COOH 10a CH₃O CH₃O CH₃ 1 none COOH 48a F F CH₃ 1 none COOH53a H F CH₃ 1 none COOH 61b F H CH₃ 2 3 COOH 62a CH₃O H CH₃ 2 none COOH61a F H CH₃ 2 none COOH 62b CH₃O H CH₃ 2 3 COOH 70a H H CH₃ 2 none COOH69b H H CH₃ 1 3 COOH 54a CF₃ H CH₃ 1 none COOH 64a CF₃ H CH₃ 2 none COOH63a OH H CH₃ 2 none COOH 63b OH H CH₃ 2 3 COOH 65a H OH CH₃ 2 none COOH65b H OH CH₃ 2 3 COOH 55b H OH CH₃ 1 3 COOH 55a H OH CH₃ 1 none COOH 56bOH H CH₃ 1 3 COOH 56a OH H CH₃ 1 none COOH 57a CH₃ H CH₃ 1 none COOH 66aCH₃ H CH₃ 2 none COOH 58a F OH CH₃ 1 none COOH 67b H CH₃O CH₃ 2 3 COOH51a Cl H CH₃ 1 none COOH 69a H H CH₃ 1 none COOH 70b H H CH₃ 2 3 COOH71a H H CH₃ 3 none COOH 68a Cl H CH₃ 2 none COOH 60a OH F CH₃ 1 noneCOOH 67a H CH₃O CH₃ 2 none COOH 59b Cl H CH₃ 2 3 COOH 69e H H H 1 — COOH70e H H H 2 — COOH 71e H H H 0 — COOH

Some examples include:

In general, in another aspect, the present invention relates todoxepin-like compounds having the formula of Formula V:

-   -   where R₉ and R₁₀ are CH₃ or connected to form a cyclopropyl        ring; R₁₃ and R₁₄ are independently F, CH₃, lower alkyl or        cyclic lower alkyl, or R₁₃ and R₁₄ are directly bonded to form a        spiro ring of size 3 to 7; R₁ is H, F, Cl, CF₃, CH₂CH₃,        CH(CH₂)₃, cyclopropyl, OCH₃, CH₂OCH₃; R₂, R₆ and R₇ are H, F,        Cl, Br, CH₃, OCH₃, CH₂OCH₃, CH₂OCH₂CH₃; Z is CO₂H,        CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl or

Where at least one of R₁, R₂, R₆, R₇, R₁₃, R₁₄ and at least one ofR₉-R₁₀, are not hydrogen when Z is COOH. Also, when R₁, R₆, R₇, R₁₃, andR₁₄ are each hydrogen, R₂ is not H, CH₃, CF₃, Cl or Br. When R₁, R₇,R₁₃, and R₁₄ are each hydrogen, and R₆ is CH₂OH, R₂ is not H, CH₃, CF₃,Cl or Br. When R₁, R₇, R₁₃, and R₁₄ are each hydrogen, and R₆ isCH₂CH₂OH, R₂ is not H, CH₃, CF₃, Cl or Br.

When each of R₁, R₆, R₇, R₁₃ and R₁₄ is H, R₂ is preferably F. When eachof R₁, R₇, R₁₃ and R₁₄ is H, and R₆ is OCH₃, R₂ is preferably OCH₃. Wheneach of R₁, R₇, R₁₃ and R₁₄ is H, and R₆ is F, R₂ is preferably OCH₃.

In some embodiments, R₅ and R₁₀ are each methyl. In some embodiments, R₉and R₁₀ are each ethyl. In other embodiments, R₉ and R₁₀ (along with thecarbon they are attached to) are connected to form a spiro ring of size3-7. In particular embodiments, R₉ and R₁₀ (along with the carbon theyare attached to) are connected to form a three-membered spiro(cyclopropyl) ring.

Table 2 provides specific embodiments of di-substituted doxepin analogshaving the following general formulae:

Table 2 provides specific embodiments of di-substituted doxepin analogs:TABLE 2 Compound Substituent Number R₂ R₃ R₆ R₇ R₃, R₆  1a, b H OCH₃ F HR₂, R₇  2a, b F H H OCH₃ R₂, R₇  3a, b OCH₃ H H OCH₃ R₂, R₇  4a, b OCH₃H H F R₂, R₇  5a, b OCH₃ H H CH₃ R₂, R₃  6a, b F OCH₃ H H R₂, R₇  7a, bF H H CH₃ R₂, R₃  8a, b OCH₃ F H H R₂, R₆  9a, b F H CH₃ H R₂, R₆ 10a, bOCH₃ H OCH₃ H R₂, R₆ 11a, b OCH₃ H CH₃ H R₃, R₆ 12a, b H OCH₃ CH₂OCH₃ HR₂, R₆ 13a, b F H CH₂OCH₃ H R₂, R₆ 14a, b OCH₃ H CH₂OCH₃ H R₂, R₆ 15a, bCH₂OCH₃ H OCH₃ H R₂, R₆ 16a, b CH₂OCH₃ H CH₂OCH₃ H R₂, R₆ 17a, b CH₃ H FH R₂, R₆ 18a, b CH₃ H CH₃ H R₂, R₆ 19a, b CH₃ H OCH₃ H R₂, R₆ 20a, b CH₃H H F

Table 3 provides specific embodiments of tri-substituted doxepinanalogs: TABLE 3 Compound Substituent Number R₂ R₃ R₆ R₇ R₂, R₃, R₆ 21a,b F OCH₃ F H R₂, R₃, R₆ 22a, b F OCH₃ OCH₃ H R₂, R₃, R₆ 23a, b OCH₃ CH₃OCH₃ H R₂, R₃, R₆ 24a, b OCH₃ CF₃ OCH₃ H R₂, R₃, R₆ 25a, b F OCH₃CH₂OCH₃ H R₂, R₃, R₆ 26a, b OCH₃ H OCH₃ F R₂, R₃, R₆ 27a, b, OCH₃ F OCH₃H R₂, R₃, R₆ 28a, b OCH₃ H F F R₂, R₃, R₆ 29a, b OCH₃ H F OCH₃

The compounds of Table 2 and Table 3 have the following chemicalstructures:

Other compounds of the invention include the following:

In general, in another aspect, the present invention relates to the useof doxepin-analogs of Formulae I-VIc to modulate sleep. Preferably,compounds of Formulae I-VIc modulate sleep with decreased side effects:e.g., the compounds do not inhibit REM sleep (consequently, sleepinduced by these compounds may more closely resemble a person's naturalsleep cycles), use of the compounds does not result in rebound insomnia,and/or the compounds do not inhibit locomotor activity or adverselyeffect body temperature.

The preferred in vitro selection criteria for doxepin analogs of theinvention are shown in Table 4. TABLE 4 In Vitro Binding Criteria H1Binding Ki < 500 nMolar (Primary Target) Off Target Binding CholinergicM1, M2, M3 Ki > 10 times the measured H1 receptor Ki Dopamine D1, D2, D3Ki > 10 times the measured H1 receptor Ki Adrenergic α1, α2 Ki > 10times the measured H1 receptor Ki

Preferably, the off target binding Ki is 50 times the measured H1receptor Ki. In some embodiments, the off target binding Ki is 100 timesthe measured H1 receptor Ki.

In vitro binding assays are used to determine H1 binding (i.e., primarytarget binding) and M1, M2 and M3 binding (i.e., off target binding).These binding assays measure the ability of doxepin analogs to displaceknown standards from the H1, M1, M2, and M3 receptors, wherein H1 is ahistamine receptor, and M1, M2, and M3 are cholinergic (muscarinic)receptors. Similar assays are performed with H1 and dopamine receptors(D1, D2 and D3), and with H1 and adrenergic receptors (α1 and α2).

The binding studies against the histamine receptor, H1, indicate bindingaffinity, and therefore, the results of the binding assays are anindication of the activity of the a doxepin analog compound. The bindingstudies against the muscarinic receptors indicate the extent to whichthe compounds bind the muscarinic receptors responsible foranti-cholinergic activity of the compound. Binding to muscarinicreceptors results in several undesired side effects of many knownantihistamines, e.g., dry-mouth. A decrease in the binding of thecompounds to the M1-M3 receptors, relative to the binding of thecompound to the H1 receptor, is an indication of the greater specificityof the compound for the histamine receptor over the muscarinic receptor.Moreover, a drug with increased specificity for the histamine receptorpossesses less anti-cholinergic side effects.

The H1 binding of doxepin analogs of the invention (also referred toherein as “test compounds” or “compounds of the invention”) isdetermined by measuring the specific binding of a given test compound,or series of test compounds, to the H1 receptor, and comparing it withthe specific binding of known standard (i.e., reference compound).Reference compounds used in this H1 binding assay include, for example,triprolidine (K_(i) 3.3 nM), chlorphenirramine (K_(i) 103.0 nM),pyrilamine (K_(i) 1.9 nM), cyproheptadine (K_(i) 8.5 nM), cimetidine(K_(i)>10,000) and dimaprit (K_(i)>10,000). (See e.g., Chang et al., J.Neurochem., 32:1653-63 (1979) (with modifications); Martinez-Mir, etal., Brain Res., 526:322-27 (1990); and Haaksme, et al., Pharmac. Ther.,47:73-104 (1990).

For example, in one embodiment of the H1 binding assay, the H1 receptoris from bovine cellular membranes, and a radioligand, [³H]Pyrilamine(15-25 Ci/mmol) at a final ligand concentration of 2.0 nM is used todetect specific binding for the H1 receptor. The assay characteristicsinclude a K_(D) (binding affinity) of 1.3 nM and a B_(max) (receptornumber) of 6.2 fmol/mg tissue (wet weight). Tripolidine (10 μM) is usedas the non-specific determinant, reference compound and positivecontrol. Binding reactions are carried out in 50 mM NA-KPO₄ (pH 7.5) at25° C. for 60 minutes. The reaction is terminated by rapid vacuumfiltration onto glass fiber filters. The level of radioactivity trappedon the filters is measured and compared to control values to ascertainany interaction between a given test compound and the H1 binding site.

The M1 binding assay determines the M1 binding of a test compound bymeasuring the specific binding of a given test compound to M1 andcomparing it with the specific binding of a reference compound. (Seee.g., Buckley, et al., Mol. Pharmacol. 35:469-76 (1989) (withmodifications)). Reference compounds used in the M1 binding assayinclude, for example, scopolamine, MethylBr (K_(i) 0.09 nM); 4-DAMPmethiodide (K_(i) 0.27 nM); pirenzepine (K_(i) 2.60 nM); HHSID (K_(i)5.00 nM); and methoctramine (K_(i) 29.70 nM).

For example, in one embodiment of the M1 binding assay, the M1muscarinic receptor is a human recombinant M1 expressed in CHO cells,and a radioligand, [³H]-scopolamine, N-methyl chloride (80-100 Ci/mmol)at a final ligand concentration of 0.5 nM is used to detect specificbinding for M1. The assay characteristics include a K_(D) (bindingaffinity) of 0.05 nM and a B_(max) (receptor number) of 4.2 pmol/mgprotein. (−)-scopolamine, methyl-, bromide (methylscopolamine bromide)(1.0 μM) is used as the non-specific determinant, reference compound andpositive control. Binding reactions are carried out in PBS for 60minutes at 25° C. The reaction is terminated by rapid vacuum filtrationonto glass fiber filters. The level of radioactivity trapped on thefilters is measured and compared to control values to ascertain anyinteraction between a given test compound and the cloned muscarinic M1binding site.

The M2 binding assay determines the M2 binding of a test compound bymeasuring the specific binding of a given test compound to M2 andcomparing it with the specific binding of a reference compound. (Seee.g., Buckley, et al., Mol. Pharmacol. 35:469-76 (1989) (withmodifications)). Reference compounds used in the N2 binding assayinclude, for example, scopolamine, MethylBr (K_(i) 0.3 nM); 4-DAMPmethiodide (K_(i) 20.7 nM); methoctramine (K_(i) 20.460 nM); HHSID(K_(i) 212.7 nM); and pirenzepine (K_(i) 832.9 nM).

For example, in one embodiment of the M2 binding assay, the M2muscarinic receptor is a human recombinant M2 expressed in CHO cells,and a radioligand, [³H]-scopolamine, N-methyl chloride (80-100 Ci/mmol)at a final ligand concentration of 0.5 nM is used to detect specificbinding for M1. The assay characteristics include a K_(D) (bindingaffinity) of 0.29 nM and a B_(max) (receptor number) of 2.1 pmol/mgprotein. (−)-scopolamine, methyl-, bromide (methylscopolamine bromide)(1.0 μM) is used as the non-specific determinant, reference compound andpositive control. Binding reactions are carried out in PBS for 60minutes at 25° C. The reaction is terminated by rapid vacuum filtrationonto glass fiber filters. The level of radioactivity trapped on thefilters is measured and compared to control values to ascertain anyinteraction between a given test compound and the cloned muscarinic M2binding site.

The M3 binding assay determines the M3 binding of a test compound bymeasuring the specific binding of a given test compound to M3 andcomparing it with the specific binding of a reference compound. (Seee.g., Buckley, et al., Mol. Pharmacol. 35:469-76 (1989) (withmodifications)). Reference compounds used in the M3 binding assayinclude, for example, scopolamine, MethylBr (K_(i) 0.3 nM); 4-DAMPmethiodide (K_(i) 0.8 nM); HHSID (K_(i) 14.5 nM); pirenzepine (K_(i)153.3 nM); and methoctramine (K_(i) 700.0 nM).

For example, in one embodiment of the M3 binding assay, the M3muscarinic receptor is a human recombinant M3 expressed in CHO cells,and a radioligand, [³H]-scopolamine, N-methyl chloride (80-100 Ci/mmol)at a final ligand concentration of 0.2 nM is used to detect specificbinding for M1. The assay characteristics include a K_(D) (bindingaffinity) of 0.14 nM and a B_(max) (receptor number) of 4.0 pmol/mgprotein. (−)-scopolamine, methyl-, bromide (methylscopolamine bromide)(1.0 μM) is used as the non-specific determinant, reference compound andpositive control. Binding reactions are carried out in 50 mM TRIS-HCl(pH 7.4) containing 10 mM MgCl₂, 1 mM EDTA for 60 minutes at 25° C. Thereaction is terminated by rapid vacuum filtration onto glass fiberfilters. The level of radioactivity trapped on the filters is measuredand compared to control values to ascertain any interaction between agiven test compound and the cloned muscarinic M3 binding site.

Preferred in vitro selection criteria for doxepin analogs of theinvention are shown in Table 5. TABLE 5 In Vitro Binding Criteria H1Binding (Primary Target) Ki < 150 nMolar Off Target Binding CholinergicM1 Ki > 10 uM Cholinergic M2 Ki > 10 uM Cholinergic M3 Ki > 10 uM

H1 binding (primary target binding) and M1, M2 and M3 binding (offtarget binding) are determined using the H1, M1, M2 and M3 bindingassays described above.

Other in vitro selection criteria for doxepin analogs of the inventionincludes hERG binding. Primary target binding and off target binding aredetermined as described above. If the test compound exhibits the desiredprimary target (H1) binding and primary target/off target bindingration, hERG binding (off target binding) is determined using a hERGblock comparative study to evaluate the effect of a given test compoundon cloned hERG channels expressed in mammalian cells. (See e.g., Brownand Rampe, Pharmaceutical News 7:15-20 (2000); Rampe et al., FEBS Lett.,417:28-32 (1997); Weirich and Antoni, Basic Res. Cardiol. 93 Suppl.1:125-32 (1998); and Yap and Camm, Clin. Exp. Allergy, 29 Suppl 3,174-81 (1999)).

Off target binding of hERG, the cardiac potassium channel responsiblefor the rapid delayed rectifier current (I_(Kr)) in human ventricles, isevaluated because inhibition of I_(Kr) is the most common cause ofcardiac action potential prolongation by non-cardiac drugs. (See Brownand Rampe (2000), Weirich and Antoni (1998); and Yap and Camm (1999)).Increased action potential duration causes prolongation of the QTinterval that has been associated with a dangerous ventriculararrhythmia, torsade de pointes. (Brown and Rampe (2000)).

In the hERG assay, hERG channels are expressed in a human embryonickidney cell line (HEK293) that lacks endogenous I_(Kr). Expression in amammalian cell line is preferable to transient expression in Xenopusoocytes, as the latter demonstrates a consistent 10-100 fold lowersensitivity to hERG channel blockers. (See, Rampe 1997).

In one embodiment of the hERG assay, the positive control (i.e.,reference compound) is terfenadine (Sigma, St. Louis Mo.), which hasbeen shown, at a concentration of 60 nM, to block hERG current byapproximately 75%. Test compounds are delivered in HEPES-bufferedphysiological saline (HB-PS)+0.1% dimethyl sulfoxide (DMSO). Each testcompound is applied at a concentration of 10 μM to the HEK293 cellsexpressing hERG (n≧3, where n=the number of cells). Cells are exposed tothe test compound for the time necessary to reach steady-state block,but not longer than 10 minutes. The positive control (60 mM terfenadine)is applied to two cells (n≧2).

The hERG-exposed cells are then transferred to the recording chamber andsuperfused with HB-PS solution. The pipette solution for whole cellrecordings includes potassium aspartate (130 mM), MgCl₂ (5 mM), EGTA (5mM), ATP (4 mM), and HEPES (10 mM) at a pH adjusted to 7.2 with KOH.Onset and steady state block of hERG current due to the test compoundare measured using a pulse pattern with fixed amplitudes(depolarization: +20 mV for 2 seconds; repolarization: −50 mV for 2seconds), repeated at 10 second intervals, from a holding potential of−80 mV. Peak tail current is measured during the 2 second step to −50mV. A steady state is maintained for at least 30 seconds before applyingthe test compound or positive control compound. Peak tail currents aremeasured until a new steady state is achieved.

Typical hERG current tracings recorded at 22° C. for a vehicle controland a positive control are shown in FIG. 1. Superimposed records incontrol and after application of a test compound. The lower panel showsvoltage stimulus (prepulse +20 mV; test pulse, −50 mV; holdingpotential, −80 mV).

In addition to the preferred, most preferred and the other in vitroselection criteria described above, doxepin analogs of the invention areselected using the following preferred in vivo sleep-wake andphysiological assessments:

NonREM Sleep: Doxepin analogs are selected if, in adult, male Wistarrats, (i) peak nonREM amount exceeds 55% nonREM per hour by no laterthan the third hour post-treatment; and (ii) the nature of this increasein nonREM sleep is such that the net cumulative total increase in nonREMsleep in the initial 6 hours post-treatment (adjusted for baseline atthe corresponding circadian time 24 hours earlier, and relative toVehicle control treatment) is not less than 20 minutes in total forcompound doses that produces maximum sleep consolidation as measured bysleep bout length, when drug is delivered orally.

The term “nonREM peak sleep time” is defined as an absolute peak amountof nonREM sleep per hour post treatment, with drug administrationoccurring at Circadian Time (CT) 18, which is 6 hours after lights offin a nocturnal laboratory rat when housed in a LD 12:12 (12-hours lightand 12 hours dark) light-dark cycle. The nominal criteria of 55% nonREMsleep per hour is equivalent to 33 minutes of nonREM sleep per hour.

As used herein, the term “cumulative nonREM sleep” is defined as the nettotal aggregate increase in the number of minutes of nonREM sleep,measured through out the entire duration of a drug's soporific effect,which typically, but not always occurs in the first 6 hourspost-treatment, adjusted for the net total aggregate number of minutesof nonREM sleep that occurred during the corresponding non-treatmentbaseline times of day recorded 24 hours earlier, relative to likevehicle control treatment.

As defined herein, the term “sleep bout” refers to a discrete episode ofcontinuous or near continuous sleep, comprised of nonREM sleep, REMsleep, or both nonREM and REM sleep stages, delimited prior and afterthe episode by greater than two contiguous 10 second epochs ofwakefulness. The following non-limiting description illustrates thisconcept: WWWWSSSSWSSSSSSSWWSSSSSSSWWWW, wherein each letter representsthe predominant state of arousal (S=sleep, W=wake) observed each 10seconds. The measured sleep “bout” is 21 ten-second epochs or 3.5minutes in duration.

Sleep Consolidation: Doxepin analogs are selected if, in adult maleWistar rats, (i) the absolute duration of longest continuous sleepepisodes (i.e., “sleep bout”) post-treatment is greater than 13 minutesin duration; (ii) the net longest sleep bout post treatment is greaterthan or equal to 3 minutes when adjusted for baseline 24 hours earlierand calculated relative to vehicle treatment; and (iii) the meanabsolute duration of every sleep bout when averaged per hour, on an hourby hour basis, is greater than or equal to 5 minutes. The aforementionedselection criteria assume that stages of sleep and wakefulness aredetermined continuously every 10 seconds (e.g., 10 second sleep scoring“epochs”), that sleep and wakefulness are measured polygraphically usingEEG and EMG criteria, and sleep episodes (comprised of nonREM and/or REMsleep) are defined as continuous “bouts” until the episode isinterrupted by greater than two contiguous 10 second epochs ofwakefulness.

As used herein, the term “longest sleep bout length” is defined as thetotal number of minutes an animal remains asleep (nonREM and/or REMsleep stages) during the single longest sleep bout that occurredbeginning in a given hour post-treatment. The “sleep bout length”measurement criteria assumes sleep is measured continuously in 10 secondepochs, and is scored based upon the predominant state, computed orotherwise determined as a discrete sleep stage (where sleep stages aredefined as nonREM sleep, REM sleep, or wakefulness) during the 10 secondinterval that defines the epoch.

The term “average sleep bout length” is defined as the average duration(in minutes) of every and all sleep episodes or bouts that began in agiven hour, independent of the individual duration of each episode orbout.

Concurrently Measured Side Effects: Doxepin analogs are selected if, inadult, male Wistar rats, these compounds (i) do not produce appreciableamounts of rebound insomnia; (ii) do not appreciably inhibit REM sleep;and (iii) do not disproportionately inhibit locomotor motor activityand/or motor tone relative to the normal effects of sleep itself. Thethreshold definitions for these three side-effect variables are asfollows:

“Rebound insomnia” is defined as period of rebound, paradoxical, orcompensatory wakefulness that occurs after the sleep promoting effectsof a hypnotic or soporific agent. Rebound insomnia is typically observedduring the usual circadian rest phase 6-18 hours post-treatment at CT-18(6 hours after lights-off, given LD 12:12), but can occur at any timeduring the initial 30 hours post-treatment. Rebound is consideredunacceptable when, in the adult, male Wistar rat, excess cumulativewakefulness associated with rebound insomnia is greater than 20% of thenet cumulative increase in sleep produced by the hypnotic or soporificeffects of a compound.

In adult, male Wistar rats, rebound insomnia manifests as an increase inwakefulness relative to corresponding times at baseline (24 hoursearlier) subsequent to a drug-induced sleep effect, and rebound insomniais measured cumulatively. The following non-limiting descriptionillustrates this measurement: Compound A, administered to laboratoryrats at CT-18 (6 hours after lights-off, given LD 12:12), produced a 50minute cumulative increase in sleep time (relative to baseline sleepmeasures 24 hours earlier) during the initial 6 hours post-treatment.After the sleep promoting effects of the compound subsided, the animalsdemonstrated a cumulative increase in wakefulness relative to baseline24 hours earlier. Because the soporific effect of the compound produced50 minutes of additional sleep, a subsequent cumulative increase inwakefulness (rebound insomnia) of greater than 10 minutes total would beunacceptable.

“REM sleep inhibition” is defined as the reduction of REM sleep timepost-treatment at CT-18 (6 hours after lights-off; LD 12:12) or at CT-5(5 hours after lights-on; LD 12:12). Compounds that reduce REM sleeptime by greater than 15 minutes (relative to baseline and adjusted forvehicle treatment) when administered at either CT-18 or CT-5 areconsidered unacceptable.

As defined herein, “disproportionate locomotor activity inhibition” is areduction of locomotor activity that exceeds the normal and expectedreduction in behavioral activity attributable to sleep. Logic dictatesthat if an animal is asleep, there will normally be a correspondingreduction in locomotor activity. If a hypnotic or soporific compoundreduces locomotor activity levels in excess of 20% greater than thatexplained by sleep alone, the compound is deemed unacceptable. Locomotoractivity (LMA) or motor tone may be quantified objectively using anyform of behavioral locomotor activity monitor (non-specific movements,telemetry-based activity monitoring, 3-dimensional movement detectiondevices, wheel running activity, exploratory measures, electromyographicrecording, etc.) so long as it is measured concurrently with objectivesleep-wakefulness measures in the same animal.

In one embodiment, locomotor activity within the animal's cage ismeasured using a biotelemetry device surgically implanted in theanimal's peritoneal cavity; the implantable device and associatedtelemetry receiver detects if and how much animal moves within the cage.Sleep and wakefulness is measured in 10 second epochs simultaneously.Counts of locomotor activity per unit time are divided by the concurrentamount of wakefulness per the same unit, yielding a “locomotor activityintensity” (LMAI) measure for that unit time. Hypnotic or soporificcompounds administered at CT-18 (6 hours after lights-off; LD 12:12)that decrease locomotor activity per unit time awake by greater than 20%relative to vehicle would be judged unacceptable.

In a more preferred embodiment, the doxepin analogs of the invention areselected using the in vivo sleep-wake and physiological assessmentcriteria shown in Table 6: TABLE 6 Change from baseline value relativeto vehicle SCORE-2000 Absolute Value only NonREM Peak Time >55%sleep/hour peak Not applicable Cumulative NonREM Not applicable >20minutes at ED100 for MSBL at T₁₋₆ Longest Sleep Bout >17 minutesabsolute peak  >5 minutes Average Sleep Bout  >6 minutes absolute peakNot used in SAR cuts Rebound Insomnia <20% of net NonREM sleep Notapplicable gain REM Sleep Inhibition not applicable not to exceed 15minutes, Rx at CT5 LMAI not applicable not to exceed 20% LMAI reduction

Methods for evaluating these sleep-wake and physiological assessmentcriteria are described above. The “absolute value” shown in secondcolumn of Table 6 refers to the value as determined for each testcompound, while the “change” value shown in the third column of Table 6reflects an adjusted value in which the absolute value is the differencefrom vehicle, when the vehicle values are adjusted for baseline.

In some embodiments, the longest sleep bout is greater than 13 minutesin duration. In others, it is greater than 17 minutes in duration. Insome embodiments, the net longest sleep bout post treatment is greaterthan or equal to 3 minutes in duration. In others, it is greater than orequal to 6 minutes in duration.

Other in vivo sleep-wake and physiological assessment criteria used toselect doxepin analogs of the invention include measurement of acutebody temperature and latent body temperature as a change in baselinerelative to vehicle. The acute body temperature change should not exceed−0.50° C., and the latent body temperature change should not exceed+0.50° C. at Time 1-6 hours. The acute body temperature (T₁₋₆) isadjusted for the corresponding baseline measured 24 hours earlier,relative to vehicle (the decrease from vehicle). The latent bodytemperature, measured 7-18 hours post drug treatment (T₇₋₁₈), isadjusted for the corresponding baseline measured 24 hours earlier,relative to vehicle (the decrease from vehicle).

The invention provides a method of modulating sleep by administering toa subject a therapeutically effective amount of a compound of FormulaI-VIb or VIc or a pharmaceutically effective salt thereof. The methodmodulates sleep several ways including decreasing the time to sleeponset, increasing the average sleep bout length, and increasing themaximum sleep bout length.

The compounds, or pharmaceutically acceptable salts thereof, isadministered orally, nasally, transdermally, pulmonary, inhalationally,buccally, sublingually, intraperintoneally, intravenously, rectally,intrapleurally, intrathecally and parenterally. In a preferredembodiment, the compound is administered orally. One skilled in the artwill recognize the advantages of certain routes of administration.

The method of modulating sleep by administering to a subject atherapeutically effective amount of a compound of Formula I-VIb or VIcor a pharmaceutically effective salt thereof is used to treat a varietyof sleep disorders including circadian rhythm abnormality, insomnia,parasomnia, sleep apnea syndrome, narcolepsy and hypersomnia. In apreferred embodiment, the method treats circadian rhythm abnormalitiesincluding jet lag, shift-work disorders, delayed sleep phase syndrome,advanced sleep phase syndrome and non-24 hour sleep-wake disorder. Inanother embodiment, the method treats insomnia including extrinsicinsomnia, psychophysiologic insomnia, altitude insomnia, restless legsyndrome, periodic limb movement disorder, medication-dependentinsomnia, drug-dependent insomnia, alcohol-dependent insomnia andinsomnia associated with mental disorders.

In another embodiment, the method treats parasomnias includingsomnambulism, pavor nocturnus, REM sleep behavior disorder, sleepbruxism and sleep enuresis. In yet another embodiment, the method treatssleep apnea disorder including central sleep apnea, obstructive sleepapnea and mixed sleep apnea. Additionally, the method treats other sleepdisorders such as narcolepsy or hypersomnia.

In some embodiments, a compound of Formula I-VIb or VIc is administeredas a pharmaceutically acceptable salt. One skilled in the art willrecognize the various methods for creating pharmaceutically acceptablesalts and identifying the appropriate salt. In a preferred embodiment,the compound or pharmaceutically acceptable salt thereof is included ina pharmaceutical composition.

A “subject” includes mammals, e.g., humans, companion animals (e.g.,dogs, cats, birds, and the like), farm animals (e.g., cows, sheep, pigs,horses, fowl, and the like) and laboratory animals (e.g., rats, mice,guinea pigs, birds, and the like). Most preferably, the subject ishuman.

A subject in need of treatment has a sleep disorder that can affect thesubject's ability to fall asleep and/or remain asleep, and/or results inunrefreshing sleep.

As used herein, the term “sleep disorder” includes conditions recognizedby one skilled in the art as sleep disorders, for example, conditionsknown in the art or conditions that are proposed to be sleep disordersor discovered to be sleep disorders. See, for example, Thorpy, M JInternational Classification of Sleep Disorders, Revised: Diagnostic andCoding Manual. American Sleep Disorders Association; Rochester, Minn.1997; and ICD-9-CM, International Classification of Diseases, NinthRevision, Clinical Modification, National Center for Health Statistics,Hyattsville, Md.

For example, sleep disorders can be generally classed into dyssomnias,e.g., intrinsic, extrinsic, and circadian rhythm disorders; parasomnias,e.g., arousal, sleep-wake transition, and rapid eye movement (REM)associated disorders, and other parasomnias; disorders associated withmental, neurological, and other medical disorders; and other sleepdisorders.

Intrinsic sleep disorders include, for example, psychophysiologicalinsomnia, sleep state misperception, idiopathic insomnia, narcolepsy,recurrent hypersomnia, idiopathic hypersomnia, post-traumatichypersomnia, obstructive sleep apnea syndrome, central sleep apneasyndrome, central alveolar hypoventilation syndrome, periodic limbmovement disorder, and restless legs syndrome.

Extrinsic sleep disorders include, for example, inadequate sleephygiene, environmental sleep disorder, altitude insomnia, adjustmentsleep disorder, insufficient sleep syndrome, limit-setting sleepdisorder, sleep-onset association disorder, food allergy insomnia,nocturnal eating (drinking) syndrome, hypnotic-dependent sleep disorder,stimulant-dependent sleep disorder, alcohol-dependent sleep disorder,and toxin-induced sleep disorder.

Circadian rhythm sleep disorders include, for example, time-zone change(jet lag) syndrome, shift work sleep disorder, irregular sleep-wakepattern, delayed sleep phase syndrome, advanced sleep phase syndrome,and non-24-hour sleep-wake disorder.

Arousal sleep disorders include, for example, confusional arousals,sleepwalking and sleep terrors.

Sleep-wake transition disorders include, for example, rhythmic movementdisorder, sleep starts, sleeptalking, and nocturnal leg cramps.

REM-associated sleep disorders include, for example, nightmares, sleepparalysis, impaired sleep-related penile erections, sleep-relatedpainful erections, REM sleep-related sinus arrest, and REM sleepbehavior disorder.

Other parasomnias include, for example, sleep bruxism, sleep enuresis,sleep-related abnormal swallowing syndrome, nocturnal paroxysmaldystonia, sudden unexplained nocturnal death syndrome, primary snoring,infant sleep apnea, congenital central hypoventilation syndrome, suddeninfant death syndrome, and benign neonatal sleep myoclonus.

A “sleep disorder” also arises in a subject that has other medicaldisorders, diseases, or injuries, or in a subject being treated withother medications or medical treatments, where the subject as a resulthas difficulty falling asleep and/or remaining asleep, or experiencesunrefreshing sleep, e.g., the subject experiences sleep deprivation. Forexample, some subjects have difficulty sleeping after undergoing medicaltreatment for other conditions, e.g., chemotherapy or surgery, or as aresult of pain or other effects of physical injuries.

It is well known in the art that certain medical disorders, for example,central nervous system (CNS) disorders, e.g. mental or neurologicaldisorders, e.g., anxiety, can have a sleep disorder component, e.g.,sleep deprivation. Thus, “treating a sleep disorder” also includestreating a sleep disorder component of other disorders, e.g., CNSdisorders. Further, treating the sleep disorder component of CNSdisorders can also have the beneficial effect of ameliorating othersymptoms associated with the disorder. For example, in some subjectsexperiencing anxiety coupled with sleep deprivation, treating the sleepdeprivation component also treats the anxiety component. Thus, thepresent invention also includes a method of treating such medicaldisorders.

For example, sleep disorders associated with mental disorders includepsychoses, mood disorders, anxiety disorders, panic disorder,addictions, and the like. Specific mental disorders include, forexample, depression, obsessive compulsive disorder, affectiveneurosis/disorder, depressive neurosis/disorder, anxiety neurosis;dysthymic disorder, behavior disorder, mood disorder, schizophrenia,manic depression, delirium, and alcoholism.

Sleep disorders associated with neurological disorders include, forexample, cerebral degenerative disorders, dementia, parkinsonism,Huntington's disease, Alzheimer's, fatal familial insomnia, sleeprelated epilepsy, electrical status epilepticus of sleep, andsleep-related headaches. Sleep disorders associated with other medicaldisorders include, for example, sleeping sickness, nocturnal cardiacischemia, chronic obstructive pulmonary disease, sleep-related asthma,sleep-related gastroesophageal reflux, peptic ulcer disease, andfibrositis syndrome.

In some circumstances, sleep disorders are also associated with pain,e.g., neuropathic pain associated with restless leg syndrome; migraine;hyperalgesia, fibromyalgia, pain; enhanced or exaggerated sensitivity topain, such as hyperalgesia, causalgia and allodynia; acute pain; burnpain; atypical facial pain; neuropathic pain; back pain; complexregional pain syndromes I and II; arthritic pain; sports injury pain;pain related to infection, e.g., HIV, post-polio syndrome, andpost-herpetic neuralgia; phantom limb pain; labor pain; cancer pain;post-chemotherapy pain; post-stroke pain; post-operative pain;neuralgia; conditions associated with visceral pain including irritablebowel syndrome, migraine and angina.

Other sleep disorders include, for example, short sleeper, long sleeper,subwakefulness syndrome, fragmentary myoclonus, sleep hyperhidrosis,menstrual-associated sleep disorder, pregnancy-associated sleepdisorder, terrifying hypnagogic hallucinations, sleep-related neurogenictachypnea, sleep-related laryngospasm, and sleep choking syndrome.

Insomnia is typically classed into sleep onset insomnia, where a subjecttakes more than 30 minutes to fall asleep; and sleep maintenanceinsomnia, where the subject spends more than 30 minutes awake during anexpected sleep period, or, for example, waking before the desiredwake-up time with difficulty or an inability to get back to sleep. Thedisclosed compounds are particularly effective in treating sleep onsetand sleep maintenance insomnias, insomnia resulting from circadianrhythm adjustment disorders, or insomnia resulting from CNS disorders. Apreferred embodiment is treating a subject for a circadian rhythmadjustment disorder. Another preferred embodiment is treating a subjectfor insomnia resulting from a mood disorder. In other embodiments, asubject is treated for sleep apnea, somnambulism, night terrors,restless leg syndrome, sleep onset insomnia, and sleep maintenanceinsomnia; or more preferably, sleep onset insomnia or sleep maintenanceinsomnia. The disclosed compounds are particularly effective fortreating sleep onset insomnia. The disclosed compounds are alsoparticularly effective for treating sleep maintenance insomnia.

The dosage regimen utilizing the compounds is selected in accordancewith a variety of factors including type, species, age, weight, sex andmedical condition of the patient; the severity of the condition to betreated; the route of administration; the renal and hepatic function ofthe patient; and the particular compound or salt thereof employed. Anordinarily skilled physician or veterinarian can readily determine andprescribe the effective amount of the drug required to prevent, counteror arrest the progress of the condition.

Oral dosages of the present invention, when used for the indicatedeffects, will range between about 0.05 to 5000 mg/day orally. Effectiveamounts of the disclosed compounds typically range between about 0.01mg/kg per day and about 100 mg/kg per day, and preferably between 0.1mg/kg per day and about 10 mg/kg/day. Techniques for administration ofthe disclosed compounds of the invention can be found in Remington: theScience and Practice of Pharmacy, 19^(th) edition, Mack Publishing Co.,Easton, Pa. (1995).

For example, in some embodiments, an acid salt of a compound containingan amine or other basic group is obtained by reacting the compound witha suitable organic or inorganic acid, such as hydrogen chloride,hydrogen bromide, acetic acid, perchloric acid and the like. Compoundswith a quaternary ammonium group also contain a counter anion such aschloride, bromide, iodide, acetate, perchlorate and the like. Otherexamples of such salts include hydrochlorides, hydrobromides, sulfates,methanesulfonates, nitrates, maleates, acetates, citrates, fumarates,tartrates (e.g. (+)-tartrates, (−)-tartrates or mixtures thereofincluding racemic mixtures), succinates, benzoates and salts with aminoacids such as glutamic acid.

Salts of compounds containing a carboxylic acid or other acidicfunctional group are prepared by reacting with a suitable base. Such apharmaceutically acceptable salt is made with a base which affords apharmaceutically acceptable cation, which includes alkali metal salts(especially sodium and potassium), alkaline earth metal salts(especially calcium and magnesium), aluminum salts and ammonium salts,as well as salts made from physiologically acceptable organic bases suchas trimethylamine, triethylamine, morpholine, pyridine, piperidine,picoline, dicyclohexylamine, N,N′-dibenzylethylenediamine,2-hydroxyethylamine, bis-(2-hydroxyethyl)amine,tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine,N-benzyl-∃-phenethylamine, dehydroabietylamine,N,N′-bisdehydroabietylamine, glucamine, N-methylglucamine, collidine,quinine, quinoline, and basic amino acid such as lysine and arginine.

In some embodiments, certain compounds and their salts also exist in theform of solvates, for example hydrates, and the present inventionincludes each solvate and mixtures thereof.

The disclosed compounds, and salts or solvates thereof may exist in morethan one crystal form, e.g., as “crystal polymorphs” or “polymorphs”.Crystal polymorphs of the disclosed compounds are prepared bycrystallization under different conditions. For example, using differentsolvents or different solvent mixtures for recrystallization;crystallization at different temperatures; various modes of cooling,ranging from very fast to very slow cooling during crystallization, andthe like. Polymorphs are also obtained by heating or melting thedisclosed compounds followed by gradual or fast cooling. The presence ofpolymorphs is determined by solid probe nuclear magnetic resonancespectroscopy, infrared spectroscopy, differential scanning calorimetry,powder X-ray diffraction, and other techniques known to one skilled inthe art.

In an embodiment, the compounds described herein, and thepharmaceutically acceptable salts thereof are used in pharmaceuticalpreparations in combination with a pharmaceutically acceptable carrieror diluent. Suitable pharmaceutically acceptable carriers include inertsolid fillers or diluents and sterile aqueous or organic solutions. Thecompounds will be present in such pharmaceutical compositions in amountssufficient to provide the desired dosage amount in the range describedherein. Techniques for formulation and administration of the disclosedcompounds of the invention can be found in Remington: the Science andPractice of Pharmacy, above.

Typically, the compound is prepared for oral administration, wherein thedisclosed compounds or salts thereof are combined with a suitable solidor liquid carrier or diluent to form capsules, tablets, pills, powders,syrups, solutions, suspensions and the like.

The tablets, pills, capsules, and the like contain from about 1 to about99 weight percent of the active ingredient and a binder such as gumtragacanth, acacias, corn starch or gelatin; excipients such asdicalcium phosphate; a disintegrating agent such as corn starch, potatostarch or alginic acid; a lubricant such as magnesium stearate; and/or asweetening agent such as sucrose, lactose, saccharin, xylitol, and thelike. When a dosage unit form is a capsule, it often contains, inaddition to materials of the above type, a liquid carrier such as afatty oil.

In some embodiments, various other materials are present as coatings orto modify the physical form of the dosage unit. For instance, in someembodiments, tablets are coated with shellac, sugar or both. In someembodiments, a syrup or elixir contains, in addition to the activeingredient, sucrose as a sweetening agent, methyl and propylparabens aspreservatives, a dye and a flavoring such as cherry or orange flavor,and the like.

For some embodiments relating to parental administration, the disclosedcompounds, or salts, solvates, or polymorphs thereof, can be combinedwith sterile aqueous or organic media to form injectable solutions orsuspensions. Injectable compositions are preferably aqueous isotonicsolutions or suspensions. The compositions may be sterilized and/orcontain adjuvants, such as preserving, stabilizing, wetting oremulsifying agents, solution promoters, salts for regulating the osmoticpressure and/or buffers. In addition, they may also contain othertherapeutically valuable substances. The compositions are preparedaccording to conventional mixing, granulating or coating methods,respectively, and contain about 0.1 to 75%, preferably about 1 to 50%,of the active ingredient.

For example, injectable solutions are produced using solvents such assesame or peanut oil or aqueous propylene glycol, as well as aqueoussolutions of water-soluble pharmaceutically-acceptable salts of thecompounds. In some embodiments, dispersions are prepared in glycerol,liquid polyethylene glycols and mixtures thereof in oils. Under ordinaryconditions of storage and use, these preparations contain a preservativeto prevent the growth of microorganisms. The terms “parenteraladministration” and “administered parenterally” as used herein meansmodes of administration other than enteral and topical administration,usually by injection, and includes, without limitation, intravenous,intramuscular, intraarterial, intrathecal, intracapsular, intraorbital,intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous,subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal andintrasternal injection and infusion.

For rectal administration, suitable pharmaceutical compositions are, forexample, topical preparations, suppositories or enemas. Suppositoriesare advantageously prepared from fatty emulsions or suspensions. Thecompositions may be sterilized and/or contain adjuvants, such aspreserving, stabilizing, wetting or emulsifying agents, solutionpromoters, salts for regulating the osmotic pressure and/or buffers. Inaddition, they may also contain other therapeutically valuablesubstances. The compositions are prepared according to conventionalmixing, granulating or coating methods, respectively, and contain about0.1 to 75%, preferably about 1 to 50%, of the active ingredient.

In some embodiments, the compounds are formulated to deliver the activeagent by pulmonary administration, e.g., administration of an aerosolformulation containing the active agent from, for example, a manual pumpspray, nebulizer or pressurized metered-dose inhaler. In someembodiments, suitable formulations of this type also include otheragents, such as antistatic agents, to maintain the disclosed compoundsas effective aerosols.

A drug delivery device for delivering aerosols comprises a suitableaerosol canister with a metering valve containing a pharmaceuticalaerosol formulation as described and an actuator housing adapted to holdthe canister and allow for drug delivery. The canister in the drugdelivery device has a headspace representing greater than about 15% ofthe total volume of the canister. Often, the polymer intended forpulmonary administration is dissolved, suspended or emulsified in amixture of a solvent, surfactant and propellant. The mixture ismaintained under pressure in a canister that has been sealed with ametering valve.

For nasal administration, either a solid or a liquid carrier can beused. The solid carrier includes a coarse powder having particle size inthe range of, for example, from about 20 to about 500 microns and suchformulation is administered by rapid inhalation through the nasalpassages. In some embodiments where the liquid carrier is used, theformulation is administered as a nasal spray or drops and includes oilor aqueous solutions of the active ingredients.

Also contemplated are formulations that are rapidly dispersing dosageforms, also known as “flash dose” forms. In particular, some embodimentsof the present invention are formulated as compositions that releasetheir active ingredients within a short period of time, e.g., typicallyless than about five minutes, preferably less than about ninety seconds,more preferably in less than about thirty seconds and most preferably inless than about ten or fifteen seconds. Such formulations are suitablefor administration to a subject via a variety of routes, for example byinsertion into a body cavity or application to a moist body surface oropen wound.

Typically, a “flash dosage” is a solid dosage form that is administeredorally, which rapidly disperses in the mouth, and hence does not requiregreat effort in swallowing and allows the compound to be rapidlyingested or absorbed through the oral mucosal membranes. In someembodiments, suitable rapidly dispersing dosage forms are also used inother applications, including the treatment of wounds and other bodilyinsults and diseased states in which release of the medicament byexternally supplied moisture is not possible.

“Flash dose” forms are known in the art; see for example, effervescentdosage forms and quick release coatings of insoluble microparticles inU.S. Pat. Nos. 5,578,322 and 5,607,697; freeze dried foams and liquidsin U.S. Pat. Nos. 4,642,903 and 5,631,023; melt spinning of dosage formsin U.S. Pat. Nos. 4,855,326, 5,380,473 and 5,518,730; solid, free-formfabrication in U.S. Pat. No. 6,471,992; saccharide-based carrier matrixand a liquid binder in U.S. Pat. Nos. 5,587,172, 5,616,344, 6,277,406,and 5,622,719; and other forms known to the art.

The doxepin analogs of the invention are also formulated as “pulsedrelease” formulations, in which the analog is released from thepharmaceutical compositions in a series of releases (i.e., pulses). Thedoxepin analogs are also formulated as “sustained release” formulationsin which the analog is continuously released from the pharmaceuticalcomposition over a prolonged period.

Also contemplated are formulations, e.g., liquid formulations, includingcyclic or acyclic encapsulating or solvating agents, e.g.,cyclodextrins, polyethers, or polysaccharides (e.g., methylcellulose),or more preferably, polyanionic β-cyclodextrin derivatives with a sodiumsulfonate salt group separate from the lipophilic cavity by an alkylether spacer group or polysaccharides. In a preferred embodiment, theagent is methylcellulose. In another preferred embodiment, the agent isa polyanionic β-cyclodextrin derivative with a sodium sulfonate saltseparated from the lipophilic cavity by a butyl ether spacer group,e.g., CAPTISOL® (CyDex, Overland, Kans.). One skilled in the art canevaluate suitable agent/disclosed compound formulation ratios bypreparing a solution of the agent in water, e.g., a 40% by weightsolution; preparing serial dilutions, e.g. to make solutions of 20%, 10,5%, 2.5%, 0% (control), and the like; adding an excess (compared to theamount that can be solubilized by the agent) of the disclosed compound;mixing under appropriate conditions, e.g., heating, agitation,sonication, and the like; centrifuging or filtering the resultingmixtures to obtain clear solutions; and analyzing the solutions forconcentration of the disclosed compound.

In addition to the therapeutic formulations described above, a therapyincluding the compounds of the present invention optionally includes,co-administration with one or more additional therapies, e.g., drugs orphysical or behavioral treatments (e.g., light therapy, electricalstimulation, behavior modification, cognitive therapy, circadian rhythmmodification, and the like). Such a practice is referred to as“combination therapy.” The other therapy or therapies in the combinationtherapy include therapies recognized by one skilled in the art asdesirable in combination with the compound of the invention, forexample, therapies known to the art or therapies which are proposed ordiscovered in the art for treating sleep disorders or treating diseasesassociated with sleep disorders, for example, therapies for any of thesleep disorders or other conditions disclosed herein. In someembodiments the compound is administered as a combination therapywhereas it is administered as a monotherapy in other embodiments.

Typically, the compound is administered as a monotherapy.

One skilled in the art will appreciate that a therapy administered incombination with the compounds of the present invention is directed tothe same or a different disorder target as that being targeted by thecompounds of the present invention. Administration of the compound ofthe invention is first, followed by the other therapy; or alternatively,administration of the other therapy may be first. The other therapy isany known in the art to treat, prevent, or reduce the symptoms of thetargeted disorder, e.g., a sleep disorder, or other disorders, e.g.,other CNS disorders. In addition, some embodiments of the presentinvention have compounds administered in combination with other knowntherapies for the target disorder. Furthermore, the other therapyincludes any agent of benefit to the patient when administered incombination with the disclosed compound.

For example, in some embodiments where the other therapy is a drug, itis administered as a separate formulation or in the same formulation asthe compound of the invention. A compound of the invention isadministered in combination therapy with any one or more ofcommercially-available, over-the-counter or prescription medications,including, but not limited to antihistamines, antimicrobial agents,fungistatic agents, germicidal agents, hormones, antipyretic agents,antidiabetic agents, bronchodilators, antidiartheal agents,antiarrhythmic agents, coronary dilation agents, glycosides,spasmolytics, antihypertensive agents, antidepressants, antianxietyagents, other psychotherapeutic agents, steroids, corticosteroids,analgesics, cold medications, vitamins, sedatives, hypnotics,contraceptives, nonsteroidal anti-inflammatory drugs, blood glucoselowering agents, cholesterol lowering agents, anticonvulsant agents,other antiepileptic agents, immunomodulators, anticholinergics,sympatholytics, sympathomimetics, vasodilatory agents, anticoagulants,antiarrhythmics, prostaglandins having various pharmacologic activities,diuretics, sleep aids, antihistaminic agents, antineoplastic agents,oncolytic agents, antiandrogens, antimalarial agents, antileprosyagents, and various other types of drugs. See Goodman and Gilman's TheBasis of Therapeutics (Eighth Edition, Pergamon Press, Inc., USA, 1990)and The Merck Index (Eleventh Edition, Merck & Co., Inc., USA, 1989).

All publications and patent documents cited herein are incorporatedherein by reference as if each such publication or document wasspecifically and individually indicated to be incorporated herein byreference. Citation of publications and patent documents is not intendedas an admission that any is pertinent prior art, nor does it constituteany admission as to the contents or date of the same. The inventionhaving now been described by way of written description, those of skillin the art will recognize that the invention can be practiced in avariety of embodiments and that the foregoing description and examplesbelow are for purposes of illustration and not limitation of the claimsthat follow.

EXAMPLE 1 Synthesis of Doxepin Analogs

The compounds of the invention, and related derivatives, can besynthesized by methods known to one skilled in the art. Detailed methodsfor synthesizing these compounds are described below. See, also, PCTPublication No. WO 03/032912.

A mixture of THF (150 mL) and N,N,N′,N′-tetramethylethylenediamine (27.8mL, 0.1853 mol, 2.5 eq.) was cooled to −78° C. s-Butyllithium (0.2 mol)was added slowly (40 min) maintaining the temperature between −65 to−78° C. After an additional 20 min stirring, 4-chlorobenzoic acid (11.60g, 0.0741 mol, 1.0 eq.) dissolved in THF (150 mL) was added over aperiod of 60 minutes while maintaining the temperature between −65 to−78° C. After 2 h, iodomethane added, and stirring continued for 1 hour,at which time the cooling bath was removed. Water (164 mL) was addedslowly and the reaction mixture was allowed to warm to room temperature.The layers were then separated, and the aqueous layer was washed withtert-butyl methyl ether (3×100 mL), and acidified with HCl to pH 1-2.The product was subsequently collected by filtration, washed with water,and dried under vacuum at 60° C. to give compound B (10.63 g, 84.0%). ¹HNMR was consistent with the structure.

Compound B (10.62 g, 62.3 mmol, 1.0 eq.) was dissolved in methanol (200mL) and thionyl chloride (11.3 mL, 155.25 mmol, 2.5 eq.) was addedslowly. The reaction solution was refluxed for 5 h, the solvent wasremoved, and the oil was taken up in methylene chloride (200 mL). Theorganic layer was washed with H₂O (3×100 mL), dried over MgSO₄,filtered, concentrated, and dried to give compound C (10.86 g, 94.4%).The structure was confirmed by ¹H-NMR.

Compound C (10.86 g, 58.8 mmol, 1.0 eq.) was dissolved in carbontetrachloride (100 mL), and N-bromosuccinimide (15.7 g, 88.2 mmol., 1.5eq.) was added followed by benzoylperoxide (0.05 g). The mixture wasrefluxed overnight. The reaction mixture was then filtered, and thesolids were washed with dichloromethane. The combined organic filtratewas concentrated and dried to give compound D (7.1 g, 45.8%). Thestructure was confirmed by ¹H NMR.

Phenol (2.79 g, 29.63 mmol, 1.1 eq.) was dissolved in 2-butanone (75.0mL) and potassium carbonate (11.17 g, 80.82 mmol., 3.0 eq.) was added,followed by compound D (7.1 g, 26.94 mmol., 1.0 eq.) dissolved in2-butanone (75.0 mL). A catalytic amount of potassium iodide (0.05 g)was added and the mixture was refluxed overnight. The cooled reactionmixture was filtered and the solids were washed with 2-butanone. Thecombined filtrate was taken up in ethyl acetate (75 mL) and was washedwith 5% aqueous NaOH (2×50 mL), brine (40 mL), and water (50 mL). Theorganic phase was concentrated and purified on silica gel to givecompound E (9.32 g). The structure confirmed product by ¹H NMR.

A solution of NaOH (4.0 g, 3.0 eq.) in H₂O (20 mL) was added to compoundE (9.32 g, 1.0 eq.) dissolved in MeOH (50 mL), and refluxed for 45 min.After cooling, the solvent was removed, H₂O added (100 mL), and aqueouslayer (aq. Extract-1) washed with ethyl acetate. The product wasextracted into the ethyl acetate layer. The organic phase was thenwashed with water/5% NaOH (3×75 mL) (aq. Extract-2). Each of the aqueousextracts 1 and 2 (which were not combined) was acidified to pH 1-2 withHCl. The white precipitate obtained was taken up in dichloromethane(3×75 mL). After removal of the solvent and drying, aq. Extract-1 gave1.61 g solid containing some product but mostly compound 1, and aq.Extract-2 gave 5.68 g product (compound F). The structures wereconfirmed by ¹H NMR.

Compound F (6.0 g, 22.84 mmol., 1.0 eq.) was dissolved indichloromethane (75.0 mL) and trifluoroacetic anhydride (7.2 g, 34.26mmol., 1.5 eq.) was added, followed by a catalytic amount ofborontrifluoride etherate (0.4 mL). The reaction mixture was heated to40° C. for 4 h. The reaction mixture was washed with water (50 mL),saturated NaHCO₃ (2×50 mL), and water (50 mL). The organic phase wasdried over MgSO₄, filtered and concentrated. The crude product waspurified on 120 g RediSep column using gradient elution,heptane/ethylacetate to give compound G (3.69 g, 66.0%). The structurewas confirmed by ¹H NMR and LC/MS.

The ketone G, was subjected to McMurray reaction. Accordingly, titaniumchloride (4.05 mL, 36.85 mmol.) was slowly added to a mixture of zincdust (5.31 g, 81.2 mmol., 5.4 eq.) in anhydrous THF (60 mL) at 0° C. Themixture was then refluxed for 2.5 hours. N-carbethoxy-4-piperidone, (5.5mL, 36.3 mmol., 2.4 eq.) and ketone G (3.69 g, 15.12 mmol., 1.0 eq.)were dissolved in anhydrous THF (40.0 mL) and added to the titanium (O)mixture, and the reaction mixture was refluxed for 6 h. An aq. solutionof K₂CO₃ (150 mL of 10% aqueous solution) was then added and stirred for30 min. The mixture was subsequently filtered over pad of celite, andthe solids were washed with ethylacetate. The layers were separated andthe organic phase was collected, dried over MgSO₄, and concentrated togive the compound H (8.15 g, 80.0% pure by HPLC). The structure wasconfirmed by ¹H NMR and LC/MS.

Compound H was dissolved in ethanol (60.0 mL), and an aq. solution ofsodium hydroxide (10.2 g, 254.76 mmol., 12.0 eq.) in H₂O (15.0 mL) wasadded and refluxed overnight. The solids were filtered off, and thenwashed with ethanol. The filtrate was concentrated and the oily residuewas taken up in dichloromethane (155 mL) and H₂O (40 mL). The aqueouslayer was extracted with CH₂Cl₂ (3×50 mL) and combined with the organiclayer. The combined organic phase was washed with brine, dried overNaSO₄, filtered and concentrated to give 3.95 g of crude compound I. Thestructure of compound I was confirmed by H NMR and LC/MS and the crudematerial was taken to the next step without purification.

Compound I (2.0 g, 6.41 mmol., 1.0 eq.), K₂CO₃ (1.77 g, 12.82 mmol., 2.0eq.), halide (5.28 g, 32.05 mmol., 5.0 eq.) and DMF (25.0 mL) werecombined and heated to 100° C. overnight. The crude reaction mixture wasmixed with H₂O (30 mL) and CH₂Cl₂ (35 mL). The organic phase wasseparated and the aqueous phase was washed with CH₂Cl₂ (2×25 mL). Thecombined organic phase was washed with brine and concentrated. The crudematerial was purified on a silica column to give compound J (1.2 g). Thestructure was confirmed by ¹H NMR and LC/MS.

Compound I (2.0 g, 6.41 mmol, 1.0 eq.), aldehyde (1.7 g, 13 mmol, 2.0eq.) and CH₂Cl₂ (20 mL) were taken in a flask under nitrogen and cooledto 0° C. Na(OAc)₃BH (2.6 g, 12.32 mmol, 1.9 eq.) was added in controlledaliquots and stirred at 0° C. for 30 min. the reaction mixture wasallowed to reach room temperature and stirred overnight. The mixture wasthen diluted with CH₂Cl₂ (40 mL), an aq. solution of saturated NaHCO₃(30 mL) was subsequently added, and the reaction mixture was stirred for10 min. The organic phase was separated and the aq. phase was extractedwith CH₂Cl₂ (2×25 mL). The combined organic layer was dried (NaSO₄),concentrated, and the crude material was purified on a silica column togive compound K (1.72 g). The structure was confined by ¹H NMR andLC/MS.

Compound K (1.6 g, 3.76 mmol, 1 eq.) was dissolved in ethanol (40.0 mL).An aqueous solution of sodium hydroxide (2.0 g, 50 mmol., 13.0 eq.) inH₂O (9.0 mL) was added and refluxed overnight. The solids were filteredoff, and the solvents were then distilled off. The residue was taken upin H₂O (40 mL) and acidified with HCl to pH 1 and stirred for 20 min.The resulting solids were filtered, washed with heptane, and dried underhigh vacuum to give the compound 51a (1.59 g). The structure of thecompound 51a was confirmed by ¹H NMR, LC/MS and elemental analysis.

Schemes 1 through 15, shown below, depict the synthesis of severaldoxepin-like compounds of the invention, with various degrees ofsubstitution.

Table 7 shows analytical data for the sleep-inducing agents shown inTable 1. TABLE 7 Analytical Data for Certain Disclosed Sleep-InducingAgents Compound 47a Molecular Formula C25H29CIFNO4*0.1 NaCl MolecularWeight 467.80 (HCl salt) Mass spec  426.2 (M + 1) Elemental Analysis(adjusted to account for 0.1 mol % of NaCl) Element Calculated ObservedC 64.19 64.03 64.12 H 6.25 6.15 6.25 N 2.99 2.96 2.95 Compound 49bMolecular Formula C24H25ClFNO3 (HCl salt) Molecular Weight 429.92 (HClsalt) Mass spec  394.3 (M + 1 of free base) Elemental Analysis ElementCalculated Observed C 67.05 66.71 66.76 H 5.86 5.9 5.78 N 3.26 3.18 3.21Cl 8.25 8.19 8.27 Compound 49a Molecular Formula C24H27ClFNO3*0.15H2OMolecular Weight 434.64 (HCl salt) Mass Spec  396.2 (M + 1 of free base)Elemental Analysis Elemen Calculated Observed C 66.32 65.96 66.05 H 6.336.26 6.33 N 3.22 3.25 3.15 Cl 8.16 8.51 8.46 Compound 49c MolecularFormula C26H31ClFNO3*(o.75 H2O) (.01 TBME) Molecular Weight 474.38 Massspec 424.2 (M + 1 of free base) Elemental Analysis (adjusted to accountfor H2O, t-butyl methyl ether (TBME) Element Calculated Observed C 65.9666.01 65.86 H 6.93 6.67 6.56 N 2.95 3.01 2.91 Cl 7.47 7.32 7.43 Compound50b Molecular Formula C25H28NclO4*0.4 H2O, 0.2 HCl Molecular Weight456.455 Mass spec 406.2 (M + 1 of free base) Elemental Analysis(adjusted to account for HCl, H2O) Element Calculated Observed C 65.7865.85 65.89 H 6.4 6.25 6.18 N 3.07 3.00 3.06 Cl 9.32 9.21 9.15 Compound50a Molecular Formula C25H30ClNO4*0.1 H2O Molecular Weight 445.77 Massspec 409.2 (M + 1) Elemental Analysis (adjusted to account for 0.1 mol %of H2O) Element Calculated Observed C 67.36 67.19 67.13 H 6.83 6.92 6.88N 3.14 7.8 7.87 Cl 7.95 3.12 3.16

Compound 50a Molecular Formula C25H30ClNO4*0.1 H2O Molecular Weight445.77 Mass spec 409.2 (M + 1) Elemental Analysis (adjusted to accountfor 0.1 mol % of H2O) Element Calculated Observed C 67.36 67.19 67.13 H6.83 6.92 6.88 N 3.14 7.8 7.87 Cl 7.95 3.12 3.16

EXAMPLE 2 Sleep-Inducing Properties of Compounds of the Invention

Sleep in mammals can be divided into sleep occurring during periods ofrapid eye movement (REM), accompanied by substantial brain activity, andperiods of non-REM (NREM) sleep, accompanied by decreased brainactivity. Typically, a normal nighttime sleep period is occupiedprimarily by NREM sleep, and thus NREM cumulation can serve as a measureof total sleep cumulation, e.g., significantly decreased NREM can beassociated with insomnia and an accumulation of “sleep debt”, e.g., anaccumulated physiological need for sleep that tends to persist until asufficient amount of additional sleep is accumulated. Thus, an increasein NREM associated with a treatment can indicated the treatment'seffectiveness in treating insomnia.

Sleep quality can be associated with sleep continuity or sleepmaintenance. For example, a subject with sleep apnea wakes up numeroustimes during a sleep period, e.g., the subject has difficultymaintaining continuous sleep. Although such a subject can accumulate atypical nights length of sleep, e.g., 8 hours, the sleep is unrefreshingdue to the waking caused by the sleep apnea. Thus, an increase in thelongest uninterrupted sleep bout (LUSB, also known as longest sleepbout) associated with a treatment can indicate the treatment'seffectiveness in enhancing sleep continuity, and therefore in treatingsleep maintenance insomnia.

Sleep-wakefulness, locomotor activity and body temperature are monitoredin male Wistar rats treated with a test compound (i.e., doxepin analog)initially at a concentration of 10 mg/kg. Higher and lower doses areassayed for select compounds (e.g., as high as 45 mg/kg, and as low asnecessary to establish a no-effect dose). Treatments are administered atCT-18, the peak of the activity dominated period (6 hours afterlights-off), and produced soporific (sleep-inducing) effectscharacterized by increased non-REM sleep time, increased sleepcontinuity, but without evidence of REM sleep inhibition or reboundinsomnia.

Sleep-wakefulness, locomotor activity and body temperature weremonitored in vivo with certain of the disclosed sleep-inducing agents46a, 47a, 49a, 49b, 50a, 50b, and derivatives listed in Table 1. Adult,male Wistar rats (250 g at time of surgery, Charles River Laboratories,Wilmington Mass.) were anesthetized (2% isoflourane in medical gradeoxygen) and surgically prepared with a cranial implant to permit chronicelectro-encephalogram (EEG) and electromyogram (EMG) recording. Bodytemperature and locomotor activity were monitored via a miniaturetransmitter (Mini-Mitter, Bend, Oreg.) surgically placed in the abdomen.The cranial implant consisted of stainless steel screws (two frontal[+3.2 AP from bregma, ±2.0 ML] and two occipital [−6.9 AP, ±5.5 ML]) forEEG recording. Two Teflon®-coated stainless steel wires were positionedunder the nuchal trapezoid muscles for EMG recording. All leads weresoldered to a miniature connector prior to surgery, and gas sterilizedin ethylene oxide. The implant assembly was affixed to the skull withdental acrylic. A minimum of three weeks was allowed for surgicalrecovery.

Each rat was permanently housed in its own individual recording cagelocated within separate, ventilated compartments of custom-designedstainless steel cabinets. Each cage was enhanced with a filter-top riserand low-torque swivel-commutator. Food and water were available adlibitum. A 24-hr light-dark cycle (12 hours light, 12 hours dark) wasmaintained throughout the study. Animals were undisturbed for at least48 hours before and after treatments.

Sleep and wakefulness were determined using “SCORE-2000™” (Hypnion,Worcester, Mass.)—an internet-based sleep-wake and physiologicalmonitoring system. The system monitored amplified EEG (bandpass 1-30Hz), integrated EMG (bandpass 10-100 Hz), body temperature andnon-specific locomotor activity (LMA) via telemetry, and drinkingactivity, continuously and simultaneously. Arousal states wereclassified on-line as non-REM (NREM) sleep, REM sleep, wake, ortheta-dominated wake every 10 seconds. Total drinking and locomotoractivity counts, and body temperature were quantitiated and recordedeach minute, using EEG feature extraction and pattern-matchingalgorithms. From this data, the longest uninterrupted sleep bout (LUSB)was obtained. The classification algorithm used individually-taughtEEG-arousal-state templates, plus EMG criteria to differentiate REMsleep from theta-dominated wakefulness, plus behavior-dependentcontextual rules (e.g., if the animal was drinking, it is awake).Drinking and locomotor activity intensity (LMA) were recorded every 10seconds, while body temperature was recorded each minute. Locomotoractivity was detected by a telemetry receiver (Mini-Mitter) beneath thecage. Telemetry measures (LMA and body temperature) were not part of thescoring algorithm; thus, sleep-scoring and telemetry data wereindependent measures.

Compounds were administered at CT-18, the peak of the activity-dominatedperiod, sufficient time was allowed to view the time course of thetreatment effect before lights-on (6 hours post-treatment). Compoundswere suspended in sterile 0.25% or 0.5% methylcellulose (1-2 ml/kg).Treatments were administered orally as a bolus.

A parallel group study design was employed. Vehicle controls were drawnfrom a large pool (N>200): a subset of the pooled vehicle controls wasselected, based on computerized matching with the 24-hour pre-treatmentbaseline of the active treatment group.

The results of NREM and LUSB parameters were measured for the disclosedsleep-inducing agents 46a, 47a, 49a, 49b, 50a, 50b, and the derivativesin Table 1. Sleep inducing agents have greater values for NREM and LUSBat a given dose, particularly at low doses, and have increasing NREM andLUSB values with increasing dose. TABLE 8 Sleep Inducing Properties* ofCompounds # dose NREM LUSB # dose NREM LUSB # dose NREM LUSB 46a 3 12.15 48a 1 36.7 6.9 55b 10 7.7 2.5 10 36 8.2 3 50.8 11.2 55a 30 12.8 2 47a1 5.5 2.9 10 58.6 10 56b 30 −9.9 −2.9 3 32.3 5.5 30 39.4 4.4 56a 30 6.44.3 10 41.1 16.7 53a 1 14.6 5 57a 30 −11 −5.3 30 56.9 10 3 38.6 17.2 66a30 27.8 8.8 49b 3 22.5 10.4 10 51.5 14.4 58a 3 9.1 5.3 10 47.3 13.1 3066.3 11.9 10 −12.5 7.6 30 43.2 15.9 61b 3 11.3 3.6 67b 10 12.6 4.6 49a0.3 −6.6 2.8 10 34.8 14.6 30 21.1 10 1 18.8 4.3 30 57.4 9.7 51a 3 32.18.8 3 37.2 11.6 62a 10 23.2 7.4 10 60.3 18.2 10 45.3 14.1 30 25.2 9.1 3049.3 6.3 30 49.1 12.9 61a 3 13.9 3.1 69a 3 32.2 7.1 50b 3 10.7 6.9 10 249.7 10 32.8 13.1 10 24.2 8.1 30 45.8 10.1 30 40 13.5 30 51.9 14.3 62b 3010.3 2.5 70b 10 33.7 8.3 50a 3 12.7 4.8 30 32.6 12.9 30 43.2 13.8 1032.6 9.3 70a 10 13.2 3.7 71a 30 8.1 3.6 30 41.9 11.2 30 33.6 6 68a 320.9 4.3 51b 3 34.7 8.4 45 29.3 12.1 10 45.9 7.8 10 45 8.3 69b 3 14.95.9 30 50.8 11.9 30 60.4 128 10 44.3 6.2 60a 10 −5.2 3.7 52a 3 13.6 8.215 43.4 14.4 67a 30 −11.5 6.5 10 26.8 5.7 20 39.4 11.6 59b 10 42.4 12.830 37.2 10.1 30 41.6 14 30 23.1 6.1 52b 10 7.5 2.9 54a 30 28.6 6.1 69e 321.2 5.6 30 26.8 7.1 64a 30 33.5 12.3 10 28.4 9.7 10a 1 −5 3.1 63a 309.6 4.6 20 54.6 12.3 3 −8.9 1.6 63b 10 −4.8 2.1 70e 3 25.7 6.6 10 15.1 465a 10 6.2 1.5 10 27.4 6.6 30 30.9 6.8 65b 10 −5.7 3.9 30 43.3 15 71e 3034.5 4.1*dose is in mg/kg;NREM and LUSB are in minutes.

EXAMPLE 3 Irwin Screen Side Effects

The Irwin screen can provide useful information on potential sideeffects of compounds on general physiological and behavioural functions.The screen was conducted by administering the test compounds orally in0.25% aqueous methylcellulose using male Wistar rats, a frequently usedspecies in such studies and for which background data are readilyavailable.

The Irwin screen tests for numerous parameters in animals that have beenadministered the test compound. For example, the screen can include:in-cage effects, e.g., dispersion, respiratory rate, locomotor activity,restlessness, fighting, alertness, apathy, and exophthalmus; in-arenaeffects, e.g., transfer arousal, spatial locomotion, ptosis, startle,tail elevation, piloerection, touch escape, positional passivity,catalepsy, tighting reflex, visual placing, grip strength, pinna,corneal, pain response, and wire manoeuvre; parameters observed inhandling, e.g., cyanosis, cutaneous blood flow, hypothermia, body tone,pupil size, light-pupil response, lacrimation, grooming, red staining,salivation, and provoked biting; general scores e.g., fearfulness,irritability, abnormal gait, abnormal body carriage, tremors, twitches,convulsions, bizarre behaviour, writhing, vocalisation, diarrhoea,number of defaecations, number of urinations, moribund, lethality, andabnormalities detected. Further details can be found in Irwin, S;Comprehensive observational assessment: I a. A systematic, quantitativeprocedure for assessing the behavioural and physiological state of themouse. Psychopharmacologia (Berl.) 13: 222-257, 1968, the entireteachings of which are incorporated herein by reference.

Irwin screening of the disclosed sleep-inducing agents was performed byCovance (Princeton, N.J.) according to Irwin, above; Covance StandardOperating Procedure (current revision of SOP PHARM 8.10); relevantregulatory authority guidelines ICH (International Committee forHarmonization) Guideline (Topic S7A; CPMP/ICH/539/00) on SafetyPharmacology Studies for human pharmaceuticals (November 2000); and allprocedures carried out on live animals were subject to the provisions ofUnited Kingdom Law, in particular the Animals (Scientific Procedures)Act, 1986. which obliges all UK laboratories to maintain a local ethicalreview process to ensure that all animal use in the establishment iscarefully considered and justified; that proper account is taken of allpossibilities for reduction, refinement or replacement and that highstandards of accommodation and care are achieved.

All chemicals used were purchased from Colorcon, Ltd, Dartford Kent, UKunless otherwise noted and were of ACS reagent grade purity or higher.All test compound formulations were prepared on the day of dosing byCovance Harrogate Dispensary. The test compounds were formulated in0.25% aqueous methylcellulose at the highest concentration required.Lower doses were obtained by serial dilution of the highestconcentration using 0.25% aqueous methylcellulose. Dose levels areexpressed in terms of the amount of test compound administered withoutregard to purity or active content. All formulations were stored at roomtemperature (nominally 10 to 30° C.) in sealed containers and protectedfrom light.

An adequate number of male Wistar (Crl:WI(Glx/BRL/Han) BR:WH) rats wereobtained from Charles River Ltd. (Margate, Kent, United Kingdom). Therats were approximately 5 weeks of age and weighed between 150 and 170 gon arrival. The animals were housed in groups of no more than six inpolypropylene cages (33×15×13 cm) or (45×28×20 cm) with solid floors andGrade 10 woodflakes (Datesand Ltd., Cheshire, United Kingdom) asbedding. The cages were cleaned and dried before use. Aspen chew blockswere placed within the cages as a form of environmental enrichment.Routinely, holding rooms were maintained within acceptable limits fortemperature and relative humidity (nominally 19 to 25° C. and 40% to70%, respectively). These rooms were illuminated by fluorescent lightfor 12 hours out of each 24 hour cycle and designed to receive at least15 fresh air changes per hour. Diet (RM1.(E).SQC. (Special DietsServices Ltd. Witham, United Kingdom) and water from the mains tapsupply are provided ad libitum (except during handling). These wereroutinely analysed for specific constituents and were not found tocontain any biological or chemical entity which might interfere with thetest system. On arrival, all animals were examined for ill-health.Animals were acclimatised for a period of at least 5 days. During thistime, animals were identified by their cage labels. A veterinaryexamination was performed before the start of any experimentalprocedures to ensure their suitability for the study. Prior to the startof the study, animals were allocated randomly to treatment groups andindividually tail-marked as they come to hand. At the end of the study,the animals were euthanized.

Each animal received a single oral administration of vehicle or testarticle, using a constant dose of 1 mg/kg. Individual doses were basedon individual body weights, obtained on the day of dosing.

The Irwin screen parameters above were systematically assessed inaccordance with the relevant controls. In general, drug-induced changes,absent in normal animals, were scored using increasing integers with ‘0’being normal (+/−, present/absent may also be used). Parameters presentin normal animals were scored using an integer that allows for increasesand decreases to be recorded. Detailed observations were performed at30, 60, 90, 180 and 300 minutes post-dose. The animals were kept for a7-day post-dose period during which time they will be observed daily forgross signs of toxicity and mortality.

The overall results of the Irwin screen are summarized in Table 9. Itcan be seen that many compounds of the present invention have a cleanprofile with regard to the Irwin screen. TABLE 9 Side Effects in IrwinScreen Compound Irwin at _(——) mg/kg dose 46a — 47a — 49b 10, 30 clean;100 sleep effects through 10 h 49a 10 clean; 30, 100 sleep effectsthrough 6 h 50b 10, 30 clean; 100 minor side effects 50a clean 51b 10,30 clean; 100 sleep effects through 6 h 61b 10, 30 clean; 100 sleepeffects through 24 h 61a 10, 30 clean; 100 side effects through 24 h 70aclean 69b clean 67c 10, 30 clean; 100 lethality 51a 10 clean; 30 someside effects; 100 lethality 69a 10, 30 clean; 100 significant sideeffects through 24 h 70b 10, 30 clean; 100 some side effects 59b clean70e 10, 30 clean; 100 some side effects 71e 10, 30 clean; 100 many sideeffects

EXAMPLE 4 Disclosed Agents Have Reduced hERG Side Effects

The cardiac potassium channel, hERG, is responsible for the rapiddelayed rectifier current (IKr) in human ventricles. This channel hasbeen selected for evaluation because inhibition of IKr is the mostcommon cause of undesirable cardiac action potential prolongation bynon-cardiac drugs. Increased action potential duration causesprolongation of the QT interval that has been associated with adangerous ventricular arrhythmia, torsade de pointes (Brown, A M; Rampe,D. (2000). Drug-induced long QT syndrome: is hERG the root of all evil?;and Pharmaceutical News 7, 15-20; Rampe, D; Roy, M L; Dennis, A; Brown,A M. (1997), the entire teachings of which are incorporated herein byreference). hERG channels were expressed in a human embryonic kidney(HEK293) cell line that lacks endogenous IKr. Expression in a mammaliancell line is preferable to transient expression in Xenopus oocytesbecause the latter shows a consistent 10-100 fold lower sensitivity tohERG channel blockers. See also, for example: A mechanism for thepro-arrhythmic effects of cisapride (Propulsid): high affinity blockadeof the human cardiac potassium channel hERG. FEBS Lett. 417,28-32;Weirich, J; Antoni, H. (1998); Rate-dependence of anti-arrhythmic andpro-arrhythmic properties of class I and class III anti-arrhythmicdrugs. Basic Res Cardiol 93 Suppl 1, 125-132; and Yap, Y G; Camm, A J.(1999); and Arrhythmogenic mechanisms of non-sedating antihistamines.Clin. Exp. Allergy 29 Suppl 3, 174-181. The entire teachings of thepreceding articles are incorporated herein by reference.

The in vitro effects of the disclosed sleep-inducing agents on the hERG(human ether-à-go-go-related gene) channel current (IKr, the rapidlyactivating, delayed rectifier cardiac potassium current) were determinedby ChanTest (Cleveland, Ohio) according to Standard Operating Proceduresof ChanTest.

All chemicals used were purchased from Sigma (St. Louis, Mo.) unlessotherwise noted and were of ACS reagent grade purity or higher. Stocksolutions of test articles and terfenadine (positive control) wereprepared using dimethyl sulfoxide (DMSO) and stored frozen. Test articleand positive control concentrations were prepared by diluting stocksolutions into a HEPES(N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid])-bufferedphysiological saline (HB-PS) solution (composition in mM): NaCl, 137;KCl, 4.0; CaCl₂, 1.8; Mg Cl₂, 1; HEPES, 10; Glucose, 10; pH adjusted to7.4 with NaOH (prepared weekly and refrigerated until use). Sinceprevious results have shown that 0.3% DMSO does not affect channelcurrent, all test and control solutions will contain 0.1% DMSO. If thefinal DMSO concentration must be greater than 0.3%, to reach a specifiedtest article concentration, a separate vehicle control test with an n>2was performed at the highest final DMSO concentration. Test and controlsolutions were prepared from stock solutions on a daily basis.

Cells used were human embryonic epithelial kidney cells (HEK293; sourcestrain, American Type Culture Collection, Manassas, Va.; sub-strain,ChanTest, Cleveland, Ohio), transformed with adenovirus 5 DNA andtransfected with hERG cDNA. Stable transfectants were selected bycoexpression with the G418-resistance gene incorporated into theexpression plasmid. Selection pressure was maintained by including G418in the culture medium. Cells were cultured in Dulbecco's Modified EagleMedium/Nutrient Mixture F-12 (D-MEM/F-12) supplemented with 10% fetalbovine serum, 100 U/mL penicillin G sodium, 100 μg/mL streptomycinsulfate and 500 μg/mL G418.

Data acquisition and analyses were performed using the suite of pCLAMPprograms (Axon Instruments, CA). Steady state was a limiting constantrate of change with time (linear time dependence) before and after testarticle application. The decrease in current amplitude upon reachingsteady state was used to calculate the percent block relative tocontrol.

All experiments were performed at room temperature (18° C.-24° C.). Eachcell acted as its own control. One concentration (10 μM) of each testarticle was applied to cells expressing hERG (n≧3, where n=the numbercells). Duration of exposure to each concentration was limited to thetime necessary to reach steady-state block, but no longer than 10minutes. One concentration of the positive control article (60 nMterfenadine) was applied to two cells (n≧2). Cells were transferred tothe recording chamber and superfused with HB-PS solution. Pipettesolution for whole cell recordings were (composition in mM): potassiumaspartate, 130; Mg Cl₂, 5; EGTA (ethylene glycol tetraacetate), 5; ATP(adenosine triphosphate), 4; HEPES, 10; pH adjusted to 7.2 with KOH.Pipette solution was prepared in batches, aliquoted, stored frozen, anda fresh aliquot thawed each day. Patch pipettes were made from glasscapillary tubing using a P-97 micropipette puller (Sutter Instruments,CA). A commercial patch clamp amplifier was used for whole cellrecordings. Before digitization, current records were low-pass filteredat one-fifth of the sampling frequency.

Onset and steady state block of hERG current due to test article weremeasured using a pulse pattern with fixed amplitudes (depolarization:+20 mV for 2 s; repolarization: −50 mV for 2 s) repeated at 10 sintervals, from a holding potential of −80 mV. Peak tail current wasmeasured during the 2 s step to −50 mV. A steady state was maintainedfor at least 30 seconds before applying test article or positivecontrol. Peak tail currents were measured until a new steady state wasachieved.

Table 10 shows the % blocking of the hERG channel at the indicatedconcentrations for various disclosed sleep inducing agents. Typically,values of about 10% or less are regarded as desirable, values from about12% to about 30% can be acceptable if the compound has strongsleep-inducing performance and no other significant side effects; andvalues greater than about 30% are regarded as undesirable. TABLE 10 hERGBlocking Compound hERG at _(—) micromolar 47a   1% @ 10 49b  3.1% @ 1049a 10.9 @ 10 49c 19.7% @ 10 50b 27.1% @ 10 50a  9.7% @ 10 51b   23% @10 61b 85.9% @10 62a 13.6% @ 10 61a 12.6% @10 70a  5.1% @ 1 69b 12.6%@10 51a 40.6% 69a  6.1% @ 1 70b 50.2% 68a 25.3% @ 10 59b   28% @ 10 69e−1.4% @1 70e  2.7% @ 1 71e  3.6% @ 1

EXAMPLE 5 Disclosed Agents Have Specificity for H1 Histamine Receptors

Binding assays were performed using the disclosed sleep-inducing agents46a, 47a, 49a, 49b, 50a, 50b, and 49c, and derivatives selected fromthose listed in Table 1 in competitive binding assays with knownstandards for the H1 histamine receptor, and the M1, M2, and M3muscarinic receptors.

The histamine H1 assays are described in Chang, et al. Heterogeneity ofHistamine H₁-Receptors: Species Variation in [³H]Mepyramine Binding ofBrain Membranes. Journal of Neurochemistry. 32: 1653-1663 (1979);Martinez-Mir, M. I., Pollard, H., Moreau, J., et al. Three HistamineReceptors (H₁, H₂, and H₃) Visualized in the Brain of Human andNon-Human Primates. Brain Res. 526: 322-327 (1990); Haaksma, E. E. J.,Leurs, R. and Timmerman, H. Histamine Receptors: Subclasses and SpecificLigands. Pharmac. Ther. 47: 73-104 (1990). The muscarinic assays aredescribed in Buckley, N. J., Bonner, T. I., Buckley, C. M., and Brann,M. R. Antagonist Binding Properties of Five Cloned Muscarinic ReceptorsExpressed in CHO-K1 Cells. Mol. Pharmacol. 35: 469-476 (1989). Theassays were performed according to the preceding articles, with thefollowing modifications. Chemical reagents in the following wereobtained from Sigma, St. Louis, Mo.

For the histamine H1 assays, the receptors were obtained from bovinecerebellar membrane tissue, with a B_(max) (receptor number) of 6.2femtomol/mg tissue (wet weight) and a KD (binding affinity) of 1.3 nM. Aradioactive ligand ([³H]pyrilamine (15-25)Ci/mmol), K_(i) 1.9 nM, finalconcentration 2.0 nM) was employed, and 10 μM triprolidine (K_(i) 3.3nM) was employed as a non-specific determinant, reference compound, andpositive control. The receptor and the radioactive ligand were combinedwith the test compound at a range of test compound concentrations fromabout 10⁻¹⁰ to about 10⁻⁶ M, and the mixture was incubated out in 50 mMNa—KPO₄ (pH 7.5) at 25° C. for 60 minutes. The reaction was terminatedby rapid vacuum filtration onto glass fiber filters. Radioactivity fromthe displaced radioactive ligand trapped onto the filters was determinedand compared to control values in order to measure any interactions ofthe test compound with the histamine H1 binding site.

For the muscarinic assays, the receptors were obtained from humanrecombinant receptors expressed in CHO cells (PerkinElmer, Inc.,Wellesley, Mass.). The radioactive ligand employed was [³H]-scopolamine,N-methyl chloride (80-100 Ci/mmol). (−)-Methylscopolamine bromide, 1.0μM, was employed as the non-specific determinant, reference compound,and positive control. After incubation, reactions were terminated byrapid vacuum filtration onto glass fiber filters. Radioactivity from thedisplaced radioactive ligand trapped onto the filters was determined andcompared to control values in order to measure any interactions of thetest compound with the respective receptor.

For the M1 receptor assay, the B_(max) (receptor number) was 4.2picomol/mg protein, and the K_(D) (binding affinity) of the receptor was0.05 nM. The radioactive ligand was employed at a final concentration0.5 nM, while the (−)-methylscopolamine bromide had a K_(i) of 0.09 nM.The receptor and the radioactive ligand were combined with the testcompound at a range of test compound concentrations from about 10⁻¹² toabout 10⁻⁵ M, incubated in Dulbecco's Phosphate Buffered Saline (PBS)for 60 minutes at 25° C., and worked up as described above.

For the M2 receptor assay, the B_(max) (receptor number) was 2.1picomol/mg protein, and the K_(D) (binding affinity) of the receptor was0.29 nM. The radioactive ligand was employed at a final concentration0.5 nM, while the (−)-methylscopolamine bromide had a K_(i) of 0.3 nM.The receptor and the radioactive ligand were combined with the testcompound at a range of test compound concentrations from about 10⁻¹² toabout 10⁻⁵ M, incubated in Dulbecco's Phosphate Buffered Saline (PBS)for 60 minutes at 25° C., and worked up as described above.

For the M3 receptor assay, the B_(max) (receptor number) was 4.0picomol/mg protein, and the K_(D) (binding affinity) of the receptor was0.14 nM. The radioactive ligand was employed at a final concentration0.2 nM, while the (−)-methylscopolamine bromide had a K_(i) of 0.3 nM.The receptor and the radioactive ligand were combined with the testcompound at a range of test compound concentrations from about 10⁻¹² toabout 10⁻⁵ M, incubated in 50 mM TRIS-HCl (pH 7.4) containing 10 mMMgCl₂, 1 mM EDTA for 60 minutes at 25° C., and worked up as describedabove.

Binding to H1 can be an indication of the desired sleep-inducingactivity of the compound. Binding to muscarinic receptors showsnon-specific binding, and can indicate anti-cholinergic activity whichcan result in undesired side effects, e.g., the side effects of manyknown antihistamines, e.g., blurred vision, dry mouth, constipation,urinary problems, dizziness, anxiety, and the like. A decrease in thebinding of the compounds to the M1-M3 receptors, relative the binding ofthe compound to the H1 receptor, is an indication of the greaterspecificity of the compound for the histamine receptor over themuscarinic receptor. Moreover, a drug with increased specificity for thehistamine receptor would possess less anti-cholinergic side effects.Table 11 shows the inhibition constant K_(i) in nM for H1 and themuscarinic receptors. It can be seen that the disclosed compounds arehighly specific for H1 over the muscarinic receptors. Thus, thedisclosed compounds can be expected to exhibit good sleep-inducingperformance with limited side effects associated with muscarinicreceptor inhibition. TABLE 11 Disclosed Agents Have Specificity for H1Histamine Receptors Ki, nanomolar Compound H1 M1 M2 M3 46a 134 >10K2450 >10K 47a 111 >10K >10K >10K 49b 61.4 >10K >10K >10K 49a69.5 >10K >10K >10K 49c 29.9 >10K >10K >10K 50b 69.8 >10K >10K >10K 50a27.2 >10K >10K >10K 51b 72 >10K >10K >10K 61b 12.2 569 3090 >10K 62a128 >10K >10K >10K 61a 182 >10K >10K >10K 62b 56.5 >10K >10K >10K 70a47.9 >10K 3310 >10K 69b 54.9 >10K >10K >10K 67b 62.5 >10K >10K >10K 51a26.4 >10K >10K >10K 69a 109 >10K >10K >10K 70a 36.8 811 1360 >10K 59b66.7 >10K >10K >10K 70e 136 >10K >10K >10K 71e 60.7 12100  5130 >10K

EXAMPLE 6 Clinical Evaluation of Doxepin Analogs

The goal of a human clinical trial is to collect data on the effects ofmodified antihistamines. Such data includes, for example, clinical signsand symptoms from physical exam, adverse events, laboratory safety(e.g., hematology, serum clinical chemistry, urinalysis), vital signs(e.g., blood pressure, heart rate, temperature, respiratory rate), andelectrocardiogram (ECG) data.

The clinical trials are conducted as follows:

I. Subject Selection

A minimum of 18 subjects are used (2 enrollment groups of 9 subjectseach). Subject candidates fulfilling the following inclusion criteriaare eligible for participation in the study:

-   -   Healthy adult male subjects, 18-45 years of age.    -   Weighing at least 60 kg and within 15% of their ideal weights        (see Table of Desirable Weights of Adults, Metropolitan Life        Insurance Company, 1983).    -   Medically healthy subjects with clinically insignificant        screening results (e.g., laboratory profiles, medical histories,        ECGS, physical exam).

Subject candidates fulfilling one of the following exclusion criteriaare ineligible for participation in the study:

-   -   History or presence of significant cardiovascular, pulmonary,        hepatic, renal, hematologic, gastrointestinal, endocrine,        immunologic, dermatologic, neurologic, or psychiatric disease.    -   History or presence of sleep disorders.    -   History of chronic or seasonal allergies requiring treatment        with H1 receptor antagonists (i.e., terfenadine, astemizole)        within the 90 days prior to the study.    -   History or presence of alcoholism or drug abuse within the past        2 years.    -   Tobacco or nicotine use within the 90 days prior to the study.    -   Known hypersensitivity or idiosyncratic reaction to the study        drug, possible excipients of the study formulation (Captisol®;        sodium saccharin, F.C.C.;    -   glycerin, U.S.P.; orange flavor; methylcellulose 400 centipoise,        U.S.P.; opurified water), or related compounds.    -   Donation (standard donation amount or more) of blood or blood        products within 90 days prior to the study.    -   Participation in another clinical trial within 90 days prior to        the first dose.    -   History or presence of any disease, medical condition, or        surgery, which may have an effect on drug absorption,        metabolism, distribution, or excretion.    -   Weight loss or gain (±10%) within 30 days prior to the study.    -   Regular consumption of (e.g., more days than not) excessive        quantities of caffeine-containing beverages (e.g., more than 5        cups of coffee or equivalent per day) within 30 days prior to        the study.    -   Any condition that, in the opinion of the Investigator or        Sponsor makes the subject unsuitable for the study.    -   Use of any prohibited prior or concomitant medications.

Each subject who completes the study screening assessments, meets alleligibility criteria, and is accepted for the study is assigned a uniqueidentification number and receives designated doses of the modifiedantihistamine and placebo according to a randomization scheme. Therandomization scheme is available only to the clinic pharmacy staffpreparing the drug (who are not involved in the administration of thedrug) and is not made available to the subjects, analysts, or members ofthe staff responsible for the monitoring and evaluation of the adverseexperiences.

Subjects may be withdrawn from the study by the Principal Investigatorfor the following reasons:

-   -   Secondary occurrence of a major exclusion criteria.    -   To protect their health.    -   Adverse events.    -   Difficulties in blood collection.    -   To protect the integrity of the study.    -   Protocol violation.    -   Failure to comply with study directions.

The clinical report includes reasons for subject withdrawals as well asdetails relevant to withdrawal. Subjects withdrawn from the trial priorto study completion undergo all procedures scheduled for studycompletion. Subjects withdrawn due to any adverse event (whether seriousor non-serious) or clinically significant abnormal laboratory testvalues are evaluated by the Investigator, or a monitoring physician, andare treated and/or followed up until the symptoms or values return tonormal or acceptable levels, as judged by the Investigator.

II. Study Restrictions

Subjects do not take prescription or over-the-counter medication(including herbal products) during the 7 days preceding the study untilthe final sample of the final pharmacokinetic sampling period has beencollected. Additionally, consumption of foods and beverages containingthe following substances is prohibited as indicated:

-   -   Methylxanthine: 72 hours before each dosing and throughout the        period of sample collection, i.e., caffeine beverages and        equivalents (e.g., chocolate bars) are prohibited.    -   Alcohol: 72 hours before each dosing and throughout the period        of sample collection.

All medications taken during the 30 days prior to study start arerecorded. Any medications taken for chronic or seasonal allergies in the90 days prior to the study is recorded.

Pre-Study Subject Screening: The Informed Consent Form is administeredat screening. Within 14 days prior to dosing, medical history anddemographic data, including name, sex, age, race, body weight (kg),height (cm), alcohol use, and tobacco use are recorded. Each subjectreceives a physical examination including complete vital signs, 12-leadECG, and laboratory tests as specified. The laboratory tests include thefollowing:

-   -   a) Hematology including hemoglobin, MCV, red blood cell count,        hematocrit, MCHC, white blood cell count with differential        platelet count and MCH;    -   b) Serum Chemistry including bun, albumin, ALT (SGOT),        creatinine, alkaline phosphatase, glucose, total bilirubin,        creatine phosphokinase (CPK), sodium, uric acid, AST (SGOT) and        triglycerides;    -   c) Urinalysis including appearance and color, glucose, nitrite,        pH, ketones, urobilinogen, specific gravity, bilirubin,        leukocytes, protein and blood;    -   d) Additional Tests including HIV, urine drug screen, HbsAg,        cannabinoids, HCV, benzodiasepines, HCV, amphetamines, hepatitis        A (IgM), opiates, alcohol, cocaine, and continine.

Subject Management: Subjects are housed from at least 36 hours beforedosing until completion of the 24-hour postdose events. They will returnfor a follow-up visit one week following the final dose or upon earlywithdrawal.

Subjects remain semi-recumbent in bed for the first 4 hours followingdrug administration. However, should adverse events occur at any time,subjects are placed in an appropriate position or are permitted to liedown on their right side. Subjects do not engage in strenuous activityat any time during the confinement period.

Standard meals are provided on Day 1 and Day 2. On Day 1, subjects arerequired to fast for a minimum of 10 hours overnight before dosing andfor at least 4 hours thereafter. However, if the option for a previousdose in the fed state is used in Period 3 of Group 2, a standardhigh-fat meal is given 30 minutes prior to dose. In this case, thehigh-fat breakfast (i.e., approximately 50% of calories from fat)consists of two eggs fried in butter, two strips of bacon, two slices ofbuttered toast, four ounces of hash brown potatoes, and eight ounces ofwhole milk. Foods and beverages containing caffeine or equivalent (e.g.,chocolate bars) are prohibited during confinement.

Water is not permitted from 2 hours before until 2 hours after dosing.Water is allowed at all other times. Standard meals are provided atapproximately 4 and 9 hours after dosing, and at appropriate timesthereafter.

III. Drug Administration

Subjects receive the dose for each period as assigned according to therandomization schedule for dosing sequence for each dose (enrollment)group. Subjects receive the assigned dose in a glass dosing cup, andwithin each dose group, all doses, active and placebo, are administeredat the same volume to maintain the double-blind. Subjects are instructedto swallow the dose.

A total of 240 mL of water is given with dosing. A designated portion ofthe water (assigned by pharmacist based on dosing volume) is added tothe emptied dosing cup, swirled to rinse, and swallowed by the subject.This process is repeated twice and then the remainder of the water isconsumed by the subject.

The starting dose for the first human dose level is based on thetoxicity and safety profiles in the preclinical studies. The equivalentbody surface area conversion from human to rat is ⅙ (ToxicologicalHandbook, Michael J. Dereleko, CRC press, Boca Raton, Fla.). Based onNOAEL of 30 mg/kg/day for rat and body surface equivalent criteria, theequivalent dose in an individual of 60 kg is 300 mg/day (⅙×30 mg/kg/day[rat NOAEL]×60 kg). Based on NOAEL dose in rat (30 mg/kg/day), the doseof 3 mg is approximately {fraction (1/10)} of the NOAEL dose in rats.The highest dose proposed of 160 mg is also below the NOAEL in rats.

If a dose limiting toxicity (Grade 3 or 4 according to the grade scalemodified from the WHO Common Toxicity Criteria—Appendix I) deemed to berelated to the study medication is observed in any 2 of the 6 subjectsat any dose level, dose escalations are stopped, and the prior dose isconsidered the maximum tolerated dose (MTD).

If one subject at any dose level experiences a dose limiting toxicity,the Principal Investigator (in consultation with the Sponsor) decides,using good clinical judgment, whether to proceed to the next dose levelas planned, or to adjust the next dose level downward from the doseplanned. This consultation is done for all groups following the previousdose group to decide whether to proceed with planned doses or to adjustdoses downward. Additionally, the planned doses may be substituted withintermediate doses if emerging safety or tolerability issues becomeapparent (i.e., there does not have to be a Grade 3 or 4 event) from thepreceding dose that suggests the need to escalate more slowly.

Dose increments is only permitted if, in the opinion of the PrincipalInvestigator, adequate safety and tolerability have been demonstrated atthe previous lower dose. In all cases, the Principal Investigator usesgood clinical judgment to decide whether to adjust the dose or to stopthe study based on an assessment of all factors relevant to the safetyof the subjects.

The Principal Investigator reviews check-in data (e.g., physicalexamination results, vital signs, questionnaire, and clinical laboratoryresults (e.g., serum chemistry, hematology, urinalysis, and urine drugscreen) for clinically significant changes since screening or theprevious period. The Principal Investigator determines if the subjectwill be dosed or withdrawn for the study based on this review.

IV. Clinical Observation

A hematology panel, a serum chemistry panel and a urinalysis isperformed at screening, at each check-in, 24 hours following each dose,and one week following the final dose, or upon early withdrawal. Bloodsamples (approximately 7 mL) are collected from an indwellingintravenous catheter into evacuated glass tubes containing sodiumheparin predose and at 0.25, 0.5, 0.75, 1.0, 1.5, 2, 3, 4, 6, 8, 10, 12,18, and 24 hours postdose. Urine samples are collected predose andduring the 0-8 hour interval each period. Samples collected during theinterval are not pooled. Each void is considered a sample. The voidingtimes are at will, not scheduled (with the exception of the predose voidand the void at the end of the 8 hour interval).

Vital signs are measured during the screenings. When the time of vitalsigns coincides with an ECG only, the vital signs are taken 10 minutesprior to the ECG. When the time of vital signs coincides with a blooddraw or a blood draw and ECG, the vital signs are taken 10 minutes priorto the blood draw. Respirations and temperature is monitored atcheck-in, 24 hours following each dose, and one week following the finaldose, or upon early withdrawal. Single measurements of blood pressureand heart rate are taken after a minimum of 5 minutes in asemi-recumbent position. Measurements taken during study confinementwill be monitored with an AVS machine at check-in; 0 (predose); 0.25,0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, 18, and 24 hours postdose; andone week following the final dose, or upon early withdrawal. For anyheart rate measurement greater than 100 beats per minute, the heart ratewill be rechecked two minutes later. On Day 1, at approximately 24 hoursprior to dosing, 3 measurements of blood pressure and heart rate, taken2 minutes apart, are taken as described as described above.

A standard 12-lead ECG is performed for each subject at screening, onDay 1 at times coinciding with Day 1 times of 1 hours prior to dose and1, 1.5, 2, 3, 4, and 6 hours postdose; on Day 1 at 1 hour predose and 1,1.5, 2, 3, 4, 6, and 24 hours postdose; and one week following the finaldose or upon early withdrawal. Additional ECGs may be performed at othertimes if deemed necessary. All standard 12-lead ECGs are recorded for 10seconds. Timing and registration technique for ECGs is standardized forall subjects. Subjects should be lying down for at least 1 minute priorto each 12-lead ECG evaluation. The Principal Investigator evaluates PR,QRS, QT, and QTc intervals. When the time of ECGs coincides with a blooddraw, the ECG will be taken following the draw.

A physician examines each subject at screening, each check-in, 24 hoursfollowing each dose, and one week following the final dose, or uponearly withdrawal. Additional examinations are performed at other timesif deemed necessary.

Immediately before vital signs measurements 1 hour predose and at 1, 2,6, and 24 hours postdose (the vital signs are taken 10 minutes prior tothe blood draw designated at these times), subjects are presented avisual analogue scale and asked to draw a vertical mark across a 100 mmline at the point ranging between Very Sleepy and Alert/Wide Awake,which best describes their level of alertness at that time.

The subjects are instructed to inform the study physician or staff ofany adverse events or intercurrent illnesses experienced during thetrial. Additionally, a specific inquiry regarding adverse events isconducted prior to dosing, at 2, 4, 8, and 24 hours postdose, and oneweek following the final dose, or upon early withdrawal. Questions areposed in a non-specific manner so as not to bias the response.

Any subject who has any adverse event (whether serious or non-serious)or clinically significant abnormal laboratory test values is evaluatedby the Investigator, or a monitoring physician, and is treated and/orfollowed up until the symptoms or values return to normal or acceptablelevels, as judged by the Investigator. A physician, either on-site or ata nearby hospital emergency room, administers treatment of any seriousadverse events. Where appropriate, medical tests and examinations areperformed to document resolution of event(s). Outcome is classified as,e.g., resolved, improved, unchanged, worse, fatal, or unknown (lost tofollow-up).

V. Reporting

All adverse events occurring during the clinical trial are recorded.Adverse events are coded using MedDRA (version 4.1). An adverseevent/experience (AE) is any unwarranted medical occurrence in a patientor clinical investigation subject administered a pharmaceutical productthat does not necessarily have a causal relationship with this treatment(ICH/WHO). An adverse event (AE) is, therefore, any unfavorable andunintended sign, (including, for example, an abnormal laboratoryfinding), symptom, or disease temporally associated with the use of amedical product, whether or not considered related to the medicalproduct (ICH/WHO).

The Investigator reviews each event and assesses its relationship todrug treatment (i.e., unrelated, unlikely, possibly, probably, almostcertainly). Each sign or symptom reported is graded on a 3-pointseverity scale (mild, moderate, or severe) and the date and time ofonset, time relationship to drug dosing, duration, and outcome of eachevent is noted. The following definitions for rating severity are used:(1) Mild: The adverse event is easily tolerated and does not interferewith daily activity; (2) Moderate: The adverse event interferes withdaily activity, but the subject is still able to function; (3) Severe:The adverse event is incapacitating and requires medical intervention.

If any of the above adverse events are serious, special procedures arefollowed. All serious adverse events are reported to the Sponsor within24 hours and followed by written reports within 48 hours, whether or notthe serious events are deemed drug-related.

A Serious Adverse Event (SAE) is any untoward medical occurrence that,at any dose, results in death, is life-threatening, results inpermanently disability or incapacitation, requires inpatienthospitalization, prolongs inpatient hospitalization, is a congenitalanomaly, may jeopardize the subject or may require intervention toprevent one or more of the other outcomes listed above.

VI. Pharmacokinetics

The following pharmacokinetic parameters are computed from theindividual plasma concentrations of the modified antihistamine compoundusing a noncompartmental approach and appropriate validatedpharmacokinetic software (e.g., WinNonlin Professional). Concentrationvalues reported as BLQ are set to zero. If concentration data areavailable, interim calculations are done (non-QC.d data) between periodsif possible. Dose escalation does not depend on pharmacokineticcalculations.

Descriptive statistics, including mean, standard deviation, coefficientof variation, geometric mean, median, minimum and maximum are computedfor each pharmacokinetic parameter by dose group. Descriptive statisticsfor natural-log transformed AUC(0-t), AUC(0-inf), and Cmax for HY2901are provided for each dose level. In addition, mean and medianconcentration versus time graphs are provided.

Dose proportionality following study medication is explored by analyzingnatural log-transformed pharmacokinetic variables AUC(0-t), AUC(0-inf),and Cmax with a linear model including the natural log-transformed doseas covariates. Dose proportionality is concluded if the 95% confidenceinterval for the slope of the covariate includes the value of 1. Doselinearity for AUC(0-t), AUC(0-inf), and Cmax is also explored by alinear model.

VII. Assessment of Safety

A by-subject treatment-emergent adverse event data listing includingverbatim term, preferred term, treatment, severity, and relationship totreatment is provided.

The number of subjects experiencing adverse events and number of adverseevents is summarized by dose level using frequency counts.

Safety data including laboratory evaluations and vital signs assessmentsis summarized by dose level and time point of collection. Descriptivestatistics are calculated for quantitative safety data and frequencycounts are compiled for classification of qualitative safety data. Inaddition, a mean change from baseline table is provided for vital signsand a shift table describing out of normal range shifts is provided forclinical laboratory results.

ECG results are classified as normal and abnormal and summarized usingfrequency counts by dose group and time point of collection. Descriptivestatistics are calculated for PR, QRS, QT, and QTc intervals.

Changes in physical exams are described in the text of the final report.

Heart rate data are summarized by treatment group and time point usingdescriptive statistics, as will individual change from baseline values.Mean change from baseline results are used to compare active dose groupsto placebo at each time point. Data from six completed subjects per doselevel should provide 80% certainty to detect a difference of 20 beatsper minute. An interim analysis is completed following each period.

VIII. Assessment of Efficacy

VAS sedation scores are summarized by time point of collection for eachdose level using descriptive statistics.

EXAMPLE 7 Preclinical Evaluation of HY2901

Prior to human clinical testing of HY2901 (also referred to herein asCompound 49a), pre-clinical testing was performed. Pre-clinicalevaluation of HY2901 included the following tests:

i. Preclinical Absorption, Distribution, Metabolism and Excretion

HY2901 was administered to rats, dogs, and cynomolgus monkeys at a doseof 3 mg/kg orally and intravenously. Plasma samples were collected fromall species for pharmacokinetic analysis. HY2901 was rapidly and highlyabsorbed after oral administration in all species. The Tmax (in hours)was 0.25, 0.5, and 1.5 in the rat, dog, and monkey, respectively. Thehalf-life (in hours) was 1.1, 4.2, and 4.4 in the rat, dog, and monkey,respectively. HY2901 is 75% protein bound in rat plasma and 60% proteinbound in human plasma.

The brains were collected from rats after oral administration todetermine brain levels of the parent drug. When brain and free druglevels in plasma were compared in the rat, it was evident that the freedrug in plasma was in equilibrium with the brain (free drug to plasmaratio of 1). There was a clear correlation between plasma and brainlevels in the rat over a wide range of plasma concentrations.

Unchanged HY2901, as well as metabolites, has been found in the urine ofanimal species administered HY2901.

Cytochrome P450 inhibition was studied with HY2901 in vitro. HY2901 didnot inhibit the activity of CYP 1A2, 2C9, 2C19, 2D6, or 3A in acommercially available human microsomal preparation. In addition, the invitro rate of metabolism in rat, dog, monkey, and human hepatocytecultures was determined for HY2901. Although HY2901 was extensivelymetabolized by rat hepatocytes, little metabolism was observed in thehuman cultures.

ii. Cardiac Effects Focus

The primary toxicological issue studied during the clinical candidateselection phase of the project was QT interval prolongation.Historically, H1 antagonists have been associated with this effect. QTprolongation in rare instances can evolve into life-threatening cardiacarrhythmias. The best in vitro test to predict the likelihood of acompound causing QT prolongation, the hERG binding assay, was the testsystem chosen to study the potential of HY2901 to produce this effect.The human hERG channel, transfected to a stable cell line, was studiedelectrophysiologically and the percent inhibition of the channel currentwas reported. In a screening assay mode, HY2901 produced a 10%inhibition of the channel current at a test concentration of 10 nM. Toput this in perspective, Seldane used as a positive control produces100% block of the channel at 60 nM. In a subsequent study with HY2901,the IC50 of HY2901 at the hERG channel was determined to be 93 mM. Peakplasma levels of less than 1.5 mM were seen in rats at the mostefficacious dose of 3 mg/kg in the sleep assay. The results of thisstudy indicate that the likelihood of HY2901 causing QT prolongation isvery low.

To determine if HY2901 could produce any changes in QT interval, thecompound was studied in telemetered Beagle dogs. Dogs were implantedwith devices to continuously monitor ECG and arterial blood pressure.Dogs (groups of 4) were studied in a Latin square cross-over design,with each dog receiving 3 different doses and a placebo. Two studieswere conducted with doses of 0.3, 1, 3, 10, and 30 mg/kg. HY2901produced no changes in QT or corrected QT interval at any dose. However,a dose-related sinus tachycardia was observed that persisted at thehigher doses for 10-12 hours. The tachycardia persisted for 6-8 hourspostdose at 0.3 and 1 mg/kg. A ceiling effect to the heart rate increaseseemed to exist with heart rates peaking at approximately 150-160 inboth the 10 and 30 mg/kg dose groups. This effect was observed with thecompound when dosed intravenously or orally. In addition, a slightincrease in blood pressure paralleled the observed tachycardia.

iii. Acute Rat Study

The purpose of this study was to evaluate the toxicity and maximumtolerated dose (MTD) of the test articles when given via oral gavage torats. Male Crl: CD®(SD)IGS BR rats (3/group) were assigned to 5 groups.At initiation of dosing, animals were approximately 7 weeks old withbody weights ranging from 172 to 206 g. Each group received either 50,100, 150, 200, or 250 mg/kg of HY2901 once daily for 5 days. Allsurviving animals were sacrificed on Day 6. Assessment of toxicity wasbased on mortality, clinical observations, and body weight data.Findings were as follows:

One 200 mg/kg rat (B64498) died on Day 1. The remaining rats survived toscheduled sacrifice. Clinical signs for the unscheduled death includedsternal recumbency at 1 hour postdose, labored respiration, paleextremities, and coldness to touch at 2 hours postdose, and death at 4hours postdose in the 200 mg/kg rat (B64498). These signs are consistentwith expected toxic effects of the test article.

Hypoactivity was observed in all rats, with the majority exhibitingapparent sleep. In the ≦100 mg/kg rats, these occurred between 30minutes and 6 hours postdose. The timeframe of both findingsoccasionally extended in the 150 mg/kg rats, beginning 15 minutespostdose and/or lasting to 8 hours postdose.

A dose-response relationship was observed in the incidence and durationof ataxia. Ataxia occurred in one 50 mg/kg rat at two timepoints (15minutes apart) on Day 1 and at one timepoint on another day, in two 100mg/kg rats at isolated timepoints (between 45 minutes and 6 hourspostdose), and in all ≧150 mg/kg rats, in which the duration of theataxia increased for some rats as well.

Day 6 body weights of two 250 mg/kg rats slightly decreased relative toother rats given HY2901.

iv. Acute Dog Study

The purpose of this study was to evaluate the toxicity and themaximum-tolerated dose (MTD) of HY2901 when given as escalating dosesvia oral gavage to dogs. Two male purebred Beagles were assigned to thestudy. At initiation of dosing, animals were at least 6 months old withbody weights ranging from 8.0 to 10.9 kg. Dogs received dosepreparations containing HY2901 once daily for 3 days in escalating dosesof 25, 50, or 75 mg/kg, no dose on Day 4, and one dose of 40 mg/kg onDay 5. Dogs were not dosed on Day 4 due to the incidence and severity ofclinical signs of toxicity observed at 75 mg/kg.

The dogs were observed at 0.25, 0.5, 0.75, 1.0, 1.5, and 2.0 hours±5minutes and 4, 6, 8, and 24 hours±15 minutes postdose. They were weighedon Days 1 and 6.

Electrocardiograms were performed and blood pressures were taken priorto dosing and at 1, 4, and 24 hours after the 40 mg/kg dose on Day 5.

Based on the severity of the clinical signs observed the dogs were notdosed on Day 4 and dogs were given a final dose of 40 mg/kg on Day 5.This dose resulted in body tremors, mild to moderate ataxia, excessivesalivation and hypoactivity. The test article had minimal effect on bodyweight. Heart rates of dogs were unaffected by treatment at 40 mg/kg. Nosignificant changes were observed in blood pressure.

The severity and incidence of clinical observations increased withincreasing dose

The clinical signs considered most representative of the toxicityobserved included the following:

-   -   1. At 25 mg/kg: No clinical signs of toxicity.    -   2. At 50 mg/kg: Body tremors, mild to severe ataxia, and        hypoactivity.    -   3. At 75 mg/kg: Tremors (entire body, hind limbs, and/or head),        mild to moderate ataxia, hypoactivity.

Based on the range and severity of the clinical signs observed, the MTDis 75 mg/kg for HY2901.

v. 14-Day Rat Study with Recovery Study

The purpose of this study was to evaluate the toxicity of HY2901 whenadministered via oral gavage to rats for at least 14 days and to assessthe reversibility, persistence, or delayed occurrence of any effectsafter a recovery period of up to 14 days.

Male and female Crl:CD®(SD)IGS BR rats were assigned to seven groups,four main study groups and three groups for toxicokinetics. Each groupreceived dose preparations containing 0.25% methylcellulose, 400 cps in200 mM acetate buffer, or 10, 30, or 150 mg of test article/kg of bodyweight (mg/kg/day) at a dose volume of 5 mL/kg.

Assessment of toxicity was based on mortality, clinical and ophthalmicobservations, body weights, food consumption, clinical pathology, organweights, and macroscopic and microscopic findings. Blood samples werecollected for toxicokinetic evaluation.

Unscheduled deaths were largely restricted to the 150 mg/kg/day groups,and were higher in females than males. Within the toxicity groups, therewere five unscheduled deaths, four of which were in the 150 mg/kg/daygroup. Of these, two males died on Day 7. Male B66391, was sacrificedmoribund with clinical signs of hypoactivity, audible, laboredrespiration, swollen abdomen and coldness to touch. At necropsy, thegastrointestinal tract of this rat was severely distended with gas.Similar distension was found in Male B66403, which was found dead afterbeing observed with a swollen abdomen, hypoactivity, and audiblerespiration. Some gas was also found in the cecal lumen of 150 mg/kg/dayfemale B66494, which was sacrificed moribund on Day 7 after it was foundin lateral recumbency with tremors and labored respiration. Another 150mg/kg/day female B66503, was found dead on Day 12 of study afterexhibiting convulsions, ataxia, clear oral and nasal discharge, andaudible, labored respiration, with unremarkable necropsy findings. Allof these 150 mg/kg/day rats were found on microscopic examination tohave suppurative inflammation involving the nasal passages, and two alsohad acute inflammation of the larynx and/or trachea, findings thatcorrelate with the clinical and macroscopic observations and areconsidered test-article related. A control Female B66463, was found deadon Day 24 following nonspecific signs of debility, and necropsyobservations were unremarkable. Among the toxicokinetics groups, one 30mg/kg/day male was found dead on Day 1 shortly after blood collection.All other unscheduled deaths of toxicokinetic rats were of 150 mg/kg/dayfemales, of which six were found dead from Day 3 through Day 14 ofstudy. Causes of these deaths were not determined, but no gavage errorwas found.

Expected pharmacological effects of the test article, hypoactivity,ataxia, and the appearance of sleep, were commonly observed. Almost alldosed rats appeared to be asleep at the 1 hour postdose observation onsome day or days of the study. The day of onset of this clinical signwas somewhat dose-related:

-   -   10 mg/kg/day. some males appeared to be asleep on Day 1 but were        unaffected thereafter until approximately Day 12 and the females        were not affected on Day 1 but appeared to be asleep at 1 hour        postdose from approximately Day 7 through the remainder of        dosing;    -   30 mg/kg/day. several males were affected on Day 1 and then at        Day 8 or 9, while the female onset began on or about Day 7;    -   150 mg/kg/day. several rats of both genders appeared to be        asleep at 1 hour postdose at various days during Week 1. As a        generalization, this postdose observation became more common in        each affected rat as the study advanced. Hypoactivity was also        dose-related Episodes of audible respiration and, in some        isolated instances, labored respiration, were almost entirely        confined to 150 mg/kg/day rats and correlated with the nasal        passage inflammation found in unscheduled deaths at this dose.        Several of the 150 mg/kg/day rats of each sex exhibited at least        one episode of audible respiration.

The test article caused a depression in food consumption in 150mg/kg/day rats in Week 1, which reached statistical significance inmales. This led to a depression in mean body weight of the 150 mg/kg/daymales, which lasted through the recovery period, despite a compensatoryincrease in food consumption in Week 2. Similar but less pronouncedtrends were observed in 150 mg/kg/day females in Week 1 but there wereno parallel changes in mean body weight or body weight change in othergroups.

No ophthalmic lesions were observed in any of the groups. Findings inthe clinical pathology data were unrelated to treatment.

The mean weights of thymus, lung, and heart for 150 mg/kg/day males weresignificantly low relative to brain weight at terminal sacrifice. Theseorgan weight findings are considered to be test article-related,although there were no corresponding microscopic findings, and the malesfrom the group assigned to recovery sacrifice did not have low weightsof those organs. No treatment-related macroscopic observations ormicroscopic toxic effects were found in either terminal sacrifice ratsor recovery sacrifice rats.

Exposure to HY2901 generally increased as the dose level increased from10 to 150 mg/kg/day. The increases in Cmax and AUC(0-t) were, ingeneral, greater than dose proportional, except for Cmax at the 150mg/kg/day dose level on Day 14, where the opposite was true. HY2901 wasrapidly absorbed and readily eliminated in rats. Females appeared tohave either similar or higher Cmax and AUC(0-t) values compared tomales, especially after multiple dosing. Minimum accumulation of HY2901after multiple dosing in rats was observed.

In conclusion, the no-observed-adverse-effect level (NOAEL) for HY2901administered by oral gavage to rats for 14 days is 30 mg/kg/day.

vi. 14-Day Dog Study with Recovery Phase

The toxicity and the toxicokinetics of HY2901 when administered dailyvia oral gavage (Phase 1) or capsules (Phase 2) to dogs for at least 14days was determined. The reversibility, persistence, or delayedoccurrence of observable effects following a 7-day (Phase 1) or 14-day(Phase 2) recovery period was also assessed. Doses of 3, 10, 30, and 70mg/kg/day were studied. All Phase 1 and 2 dogs survived until scheduledsacrifice.

There were no treatment-related clinical observations in the 3 mg/kg/dayPhase 1 dogs. In Phase 2, treatment-related head tremors, ataxia,hypoactivity, and excessive salivation were noted at least once indifferent males; hypoactivity once in one female; and sporadic vomiting(white, foamy or containing food) were noted in females. There were noremarkable clinical observations during the Phase 1 and 2 recoveryperiods.

There were no changes in respiration rates, rectal body temperatures,body weights, food consumption, electrocardiograms, and ophthalmicexaminations attributed to HY2901 in Phase 1 and 2. Sinus tachycardia(heart rate above 190 beats/minute) was noted during Phase 2 in one maleon Day 3 (predose and 4 hours postdose), on Day 14 (2 and 4 hourspostdose), and on Day 29 (after 14-day recovery). Also three Phase 2females were noted with sinus tachycardia; one on Day 3 (4 hourspostdose) and on Day 14 (2 and 4 hours postdose), one on Day 14 (2 hourspostdose), and one on Day 29 (after 14-day recovery). No unequivocalevidence is present to suggest a direct treatment effect as evidenced byinconsistency in tachycardia observations; therefore, these findingswere considered incidental to the treatment.

Clinical pathology data were generally unremarkable and similar amongthe 0 and 3 mg/kg/day groups at the designated intervals during Phase 1,and were unremarkable for the 70 mg/kg/day dogs at all collectionintervals of Phase 2. There was no evidence in the clinical laboratorydata of an effect from the administration of HY2901.

In conclusion, gavage administration of HY2901 at 3 mg/kg/day and at 70mg/kg/day via gelatin capsule to purebred Beagles for at least 14 daysresulted in a NOAEL for HY2901 treatment of 70 mg/kg/day.

The above methods and protocols are useful in the pre-clinicalevaluation of other modified anti-histamine of the invention.

EXAMPLE 8 Binding Specificity of Compound 37 for H1 Histamine Receptorsand M1, M2, and M3 Cholinergic Receptors

Binding assays were performed using compound 37 (HYP10073) incompetitive binding assays with known standards for the H1 histaminereceptor, and the M1, M2, and M3 muscarinic receptors.

First, the binding specificity of compound 37 for a variety of receptorswas evaluated. The percentage inhibition (average; N=2) for thesereceptors is shown below in Table 12. TABLE 12 Inhibitory Effect ofCompound 37 Receptor 1.0E−6 NEUROTRANSMITTER RELATED Adenosine,Non-seletive 6.49% Adrenergic, Alpha 1, Non-selective 6.56% Adrenergic,Alpha 2, Non-selective −3.79% Adrenergic, Beta, Non-selective 6.97%Dopamine Transporter 2.14% Dopamine, Non-selective 21.10% GABA A,Agonist Site 20.18% GABA A, BD2, alpha 1 site −7.13% GABA-B 2.17%Glutamate, AMPA Site 3.59% Glutamate, Kainate Site 1.51% Glutamate, NMDAAgonist Site −5.56% Glutamate, NMDA, Glycine (Stry-insens Site) −12.50%Glycine, Strychnine-sensitive 1.15% Histamine, H1 23.13% Histamine, H20.80% Histamine, H3 5.11% Melatonin, Non-selective 11.29% Muscarinic, M1(hr) 5.11% Muscarinic, M2 (hr) 14.62% Muscarinic, Non-selective, Central12.84% Muscarinic, Non-selective, Peripheral 5.37% Nicotinic(a-Bungarotoxin Insensitive) 1.69% Norepinephrine Transporter 19.51%Opioid, Non-selective −9.30% Orphenin (hr) 3.53% Serotonin Transporter11.46% Serotonin, Non-selective 21.45% Sigma, Non-selective 13.27%STEROIDS Estrogen −4.68% Testosterone (cytosolic) 0.24% ION CHANNELSCalcium Channel, Type L (Dihydropyridine Site) 5.82% Calcium Channel,Type N 3.45% Potassium Channel, ATP-Sensitive 26.16% Potassium Channel,Ca2+ Act., VI −12.71% Potassium Channel, I[Kr] (hERG) (hr) −5.23%Sodium, Site 2 −4.12% SECOND MESSENGERS Nitric Oxide, NDS(Neuronal-Binding) 17.21% PROSTAGLANDINS Laukotriene, LTD4 −20.09%Thromboxane A2 (h) 6.55% GROWTH FACTORS/HORMONES Corticotropin ReleasingFactor, Non-selective 4.31% Oxytocin −5.82% Platelet Activating Factor,PAF −1.15% Thyrotropin Releasing Harmone, TRB −2.98% BRAIN/GUT PEPTIDESAngiotensin II, AT1 (h) 12.03% Angiotensin II, AT2 1.51% Bradykinin, BK2−3.04% Cholecystokinin, CCK1 (CCKA) 8.92% Cholecystokinin, CCK2 (CCKB)3.98% Endothelin, ET-A (h) 10.15% Endothelin, ET-B (h) −6.51% Galanin,Non-selective −5.07% Neurokinin, NK1 10.27% Neurokinin, NK2 (NKA) (hr)−4.17% Neurokinin, NK3 (NKB) −0.23% Vasoactive Intestinal Peptide,Non-selective 2.51% Vasopressin 1 11.55% ENZYMES CholineAcetyltransferase 6.51% Esterase, Acetylcholine 2.54% Glutamic AcidDecarboxylase −4.76% Oxidase, MAO-A, Peripheral −17.23% Oxidase, MAO-B,Peripheral 4.24%

Values in Table 12 are expressed as percentage inhibition of specificbinding, and these values represent the average of replicate tubes ateach of the concentrations tested. Bolded values represent inhibition of50% or greater.

The binding specificity for compound 37 for histamine H1 receptor andmuscarinic M1, M2 and M3 receptors was further evaluated using thebinding assays described above.

In particular, the in vitro H1 binding assay measured the specificbinding of compound 37 to the H1 receptor and compared the measuredspecific binding with the specific binding of known standard (ie.,reference compound), triprolidine. (See e.g., Chang et al., J.Neurochem., 32:1653-63 (1979) (with modifications); Martinez-Mir, etal., Brain Res., 526:322-27 (1990); and Haaksme, et al., Pharmac. Ther.,47:73-104 (1990)).

In these tests, the H1 receptor was a bovine cellular membrane, and aradioligand, [³H]Pyrilamine (15-25 Ci/mmol) at a final ligandconcentration of 2.0 nM was used to detect specific binding of compound37 for the H1 receptor. The assay characteristics include a K_(D)(binding affinity) of 1.3 nM and a B_(max) (receptor number) of 6.2fmol/mg tissue (wet weight). Tripolidine (10 μM) was used as thenon-specific determinant, reference compound and positive control.Binding reactions were carried out in 50 mM NA-KPO₄ (pH 7.5) at 25° C.for 60 minutes. The reaction was terminated by rapid vacuum filtrationonto glass fiber filters. The level of radioactivity trapped on thefilters was measured and compared to control values to ascertain anyinteraction between compound 37 and the H1 binding site.

The binding curves for compound 37 and triprolidine binding to the H1receptor are shown in FIG. 2. The calculated IC50 and K_(i) values fortriprolidine and compound 37 are shown below in Table 13. TABLE 13IC50/K_(i) Determination for Compound 37 IC50 K_(i) Slope Triprolidine6.92E−9 2.65E−9 −0.76 Compound 37 6.75E−8 2.58E−8 −0.99

The H1 specific binding and % inhibition for compound 37 at variousconcentrations, as determined using the in vitro H1 binding assay, areshown below in Table 14. TABLE 14 H1 Binding of Compound 37 at VaryingConcentrations (Average; N = 2) % Specific Receptor Concentration %Inhibition Binding NEUROTRANSMITTER 1.0E−9 2.99% 97.01% RELATED 1.0E−88.60% 91.40% Histamine, H1 3.0E−8 30.37% 69.53% 1.0E−7 60.82% 39.18%3.0E−7 81.03% 18.97% 1.0E−6 93.75% 6.25% 3.0E−6 93.26% 6.74% 1.0E−596.79% 3.21%

The values in Table 13 represent the average of replicate tubes at eachof the concentrations tested.

The in vitro M1 binding assay measured the specific binding of compound37 to M1 and compared this measured binding specificity with thespecific binding of the reference compound (−)-scopolamine, MeBr. (Seee.g., Buckley, et al., Mol. Pharmacol. 35:469-76 (1989) (withmodifications)).

In this assay, the M1 muscarinic receptor was a human recombinant M1expressed in CHO cells, and a radioligand, [³H]-scopolamine, N-methylchloride (80-100 Ci/mmol) at a final ligand concentration of 0.5 nM wasused to detect specific binding for M1. (−)-scopolamine, methyl bromide(methylscopolamine bromide) was used as the non-specific determinant,reference compound and positive control. The assay characteristicsinclude a K_(D) (binding affinity) of 0.05 nM and a B_(max) (receptornumber) of 4.2 pmol/mg protein (1.0 μM). Binding reactions were carriedout in PBS for 60 minutes at 25° C. The reaction was terminated by rapidvacuum filtration onto glass fiber filters. The level of radioactivitytrapped on the filters was measured and compared to control values toascertain any interaction between a given test compound and the clonedmuscarinic M1 binding site.

The binding curves for compound 37 and (−)-scopolamine, MeBr binding tothe M1 receptor are shown in FIG. 3. The calculated IC50 and K_(i)values for (−)-scopolamine, MeBr and compound 37 are shown below inTable 15. TABLE 15 IC50/K_(i) Determination for Compound 37 IC50 K_(i)Slope (−)-scopolamine, MeBr 6.08E−10 6.47E−11 −1.14 Compound 37 N/A N/AN/AN/A refers to a value >10,000

The M1 specific binding and % inhibition for compound 37 at variousconcentrations, as determined using the in vitro M1 binding assay, areshown below in Table 16. TABLE 16 M1 Binding of Compound 37 at VaryingConcentrations (Average; N = 2) % Specific Receptor Concentration %Inhibition Binding NEUROTRANSMITTER 1.0E−9 −10.93% 110.93% RELATED1.0E−8 −16.93% 116.93% Muscarinic, M1 (hr) 3.0E−8 −6.82% 106.82% 1.0E−71.61% 98.39% 3.0E−7 2.87% 97.13% 1.0E−6 −5.22% 105.22% 3.0E−6 −11.53%111.83% 1.0E−5 −7.57% 107.57%

The values in Table 13 represent the average of replicate tubes at eachof the concentrations tested.

The M2 binding assay measured the specific binding of the M2 receptorand compared the measured specific binding with the specific binding ofa given test compound to M2 and comparing it with the specific bindingof a reference compound, (−)-scopolamine, MeBr. (See e.g., Buckley, etal., Mol. Pharmacol. 35:469-76 (1989) (with modifications)).

In this assay, the M2 muscarinic receptor was a human recombinant M2expressed in CHO cells, and a radioligand, [³H]-scopolamine, N-methylchloride (80-100 Ci/mmol) at a final ligand concentration of 0.5 nM wasused to detect specific binding for M1. The assay characteristicsincluded a K_(D) (binding affinity) of 0.29 nM and a B_(max) (receptornumber) of 2.1 pmol/mg protein. (−)-scopolamine, methyl-, bromide(methylscopolamine bromide) (1.0 μM) was used as the non-specificdeterminant, reference compound and positive control. Binding reactionswere carried out in PBS for 60 minutes at 25° C. The reaction wasterminated by rapid vacuum filtration onto glass fiber filters. Thelevel of radioactivity trapped on the filters was measured and comparedto control values to ascertain any interaction between a given testcompound and the cloned muscarinic M2 binding site.

The binding curves for compound 37 and (−)-scopolamine, MeBr for the M2receptor are shown in FIG. 4. The calculated IC50 and K_(i) values for(−)-scopolamine, MeBr and compound 37 are shown below in Table 17. TABLE17 IC50/K_(i) Determination for Compound 37 IC50 K_(i) Slope(−)-scopolamine, MeBr 6.47E−10 2.64E−11 −0.89 Compound 37 N/A N/A N/AN/A refers to a value >10,000

The M2 specific binding and % inhibition for compound 37 at variousconcentrations, as determined using the in vitro M2 binding assay, areshown below in Table 18. TABLE 18 M2 Binding of Compound 37 at VaryingConcentrations (Average; N = 2) % Specific Receptor Concentration %Inhibition Binding NEUROTRANSMITTER 1.0E−9 −2.57% 102.57% RELATED 1.0E−8−14.04% 114.04% Muscarinic, M2 (hr) 3.0E−8 −12.52% 112.52% 1.0E−7−17.74% 117.74% 3.0E−7 −9.94% 108.94% 1.0E−6 −2.53% 102.53% 3.0E−6−5.10% 105.10% 1.0E−5 5.61% 94.39%

The values in Table 13 represent the average of replicate tubes at eachof the concentrations tested.

The M3 binding assay measured the specific binding of compound 37 to theM3 receptor and compared the measured specific binding with the specificbinding of a reference compound, (−)-scopolamine, MeBr. (See e.g.,Buckley, et al., Mol. Pharmacol. 35:469-76 (1989) (with modifications)).

In this assay, the M3 muscarinic receptor was a human recombinant M3expressed in CHO cells, and a radioligand, [³H]-scopolamine, N-methylchloride (80-100 Ci/mmol) at a final ligand concentration of 0.2 nM wasused to detect specific binding for M1. The assay characteristicsincluded a K_(D) (binding affinity) of 0.14 nM and a B_(max) (receptornumber) of 4.0 pmol/mg protein. (−)-scopolamine, methyl-, bromide(methylscopolamine bromide) (1.0 μM) was used as the non-specificdeterminant, reference compound and positive control. Binding reactionswere carried out in 50 mM TRIS-HCl (pH 7.4) containing 10 mM MgCl₂, 1 mMEDTA for 60 minutes at 25° C. The reaction was terminated by rapidvacuum filtration onto glass fiber filters. The level of radioactivitytrapped on the filters was measured and compared to control values toascertain any interaction between a given test compound and the clonedmuscarinic M3 binding site.

The binding curves for compound 37 and (−)-scopolamine, MeBr for the M3receptor are shown in FIG. 5. The calculated IC50 and K_(i) values for(−)-scopolamine, MeBr and compound 37 are shown below in Table 19. TABLE19 IC50/K_(i) Determination for Compound 37 IC50 K_(i) Slope(−)-scopolamine, MeBr 3.64E−10 1.32E−10 −0.55 Compound 37 N/A N/A N/AN/A refers to a value >10,000

The M3 specific binding and % inhibition for compound 37 at variousconcentrations, as determined using the in vitro M3 binding assay, areshown below in Table 20. TABLE 20 M3 Binding of Compound 37 at VaryingConcentrations (Average; N = 2) % Specific Receptor Concentration %Inhibition Binding NEUROTRANSMITTER 1.0E−9 −0.65% 100.65% RELATED 1.0E−8−8.34% 108.34% Muscarinic, M3 (hr) 3.0E−8 6.99% 93.01% 1.0E−7 6.85%93.15% 3.0E−7 5.55% 94.45% 1.0E−6 0.79% 99.21% 3.0E−6 5.41% 94.59%1.0E−5 4.27% 95.73%

The values in Table 13 represent the average of replicate tubes at eachof the concentrations tested.

EXAMPLE 9 SCORE-2000 Data for Compound 37

The effect of compound 37 (also referred to herein as HY-10073) on sleepand wakefulness in adult male Wistar rats were determined using the“SCORE-2000™” (Hypnion, Worcester, Mass.)—an internet-based sleep-wakeand physiological monitoring system described above in Example 2.

Sleep-wakefulness, locomotor activity and body temperature weremonitored in male Wistar rats treated with compound 37 at aconcentration of either 10 mg/kg PO (i.e., oral administration) or 30mg/kg PO. Treatments were administered at CT-18, the peak of theactivity dominated period (6 hours after lights-off), and producedsoporific (sleep-inducing) effects characterized by increased non-REMsleep time, increased sleep continuity, but without evidence of REMsleep inhibition or rebound insomnia.

When compound 37 was administered at 10 mg/kg PO, peak NREM sleep timewas 64% per hour. Cumulative NREM sleep time, adjusted for baseline andrelative to vehicle, increased by 53 minutes±6 minutes. The absolutelongest sleep bout post-treatment was 18.3 minutes, while the longestsleep bout (also referred to herein as “Longest Uninterrupted SleepBout” (LUSB)), adjusted for baseline and relative to vehicle, was 10.8minutes±2.7 minutes. The absolute average sleep bout was 9 minutes,while the average sleep bout length, adjusted for baseline and relativeto vehicle, increased by 5.3 minutes±1.4 minutes.

Also, administration of compound 37 at a concentration of 10 mg/kg, didnot produce any REM inhibition or rebound insomnia. Furthermore, therewas no disproportional reduction in either locomotor activity (LMA) orbody temperature.

When compound 37 was administered at 30 mg/kg PO, peak NREM sleep timewas 72% per hour. Cumulative NREM sleep time, adjusted for baseline andrelative to vehicle, increased by 62 minutes±9 minutes. The absolutelongest sleep bout post-treatment was 25 minutes, while the longestsleep bout (also referred to herein as “Longest Uninterrupted SleepBout” (LUSB)), adjusted for baseline and relative to vehicle, was 17.1minutes±3.6 minutes. The absolute average sleep bout was 9 minutes,while the average sleep bout length, adjusted for baseline and relativeto vehicle, increased by 5.6 minutes±1.5 minutes.

Also, administration of compound 37 at a concentration of 30 mg/kg, didnot produce any appreciable REM inhibition at CT-18. No rebound insomniawas observed with the 30 mg/kg administration. Furthermore, there was nodisproportional reduction in LMA. The drop in body temperature occurredonly at this high dose of compound 37 (i.e., an unacceptable drop inbody temperature was only observed when compound 37 was administered atsuch high dosages).

The sleep consolidating effects of compound 37 administered at aconcentration of 30 mg/kg (PO) at CT-18 is shown in FIG. 6. (CT-18 isindicated as a triangle on the x-axis of FIG. 6). As seen in FIG. 6, arobust increase in the increase in the peak sleep bout duration wasobserved in the initial 2-3 hours post treatment.

EXAMPLE 10 Evaluation of the Analgesic Activity of HY-2901 (Compound49a)

The objective of this example was to determine the analgesic activity ofHY-2901 (Also referred to herein as Compound 49a) following oraladministration. Analgesic activity was assessed abdominal spasm test inthe rat and mouse. Analgesic activity is also assessed using the tailclip test in the mouse, tail flick test in the rat, Randall-Selitto testin the rat and comparisons are made with a vehicle control group.Reference compounds ASA (acetylsalicylic acid) and morphine are alsoincluded for comparison.

The tail clip and tail flick test provide useful information about thecentral analgesic activity of the test article. The Randall-Selitto testprovides information on the compound's ability to modify a hyperalgesicstate and the abdominal spasm test provides information on theperipheral analgesic activity of the test article. The test article isadministered by oral gavage, this being the intended clinical route ofadministration. The dose levels employed are expected to encompass theefficacy dose and provide an adequate safety margin.

Test Article, Reference Compound and Irritant Formulation

All formulations were prepared on each day of dosing. The test article(HY-2901) was formulated in 0.25% (w/v) MC at the highest concentrationrequired. Lower doses were obtained by serial dilution of the highestconcentration using 0.25% (w/v) MC. The reference compound,acetylsalicylic acid, was formulated in 0.25% (w/v) MC at the requiredconcentrations. Brewer's yeast was formulated in water for injection atthe required concentration. Acetic acid was diluted with water forinjection to provide the required concentration for administration.

Dose levels will be expressed in terms of the amount of test article Ireference compound/irritant administered without regard to purity oractive content.

Animals

An adequate number of male Crl:CD-I(ICR)BR mice and Wistar rats wereobtained from Charles River (UK) Ltd., Margate, Kent. The mice wereapproximately 4 weeks of age and weigh between 18 and 22 g on arrival.The rats were approximately 5 weeks of age and weigh between 150 and 170g on arrival. The age and weight of the animals at the start of thestudy was documented in the raw data and final report,

The animals were housed in groups appropriate to the size of cagingused, in cages that conform to the Code of Practice for the housing andcare of animals used in the Scientific Procedures Act (Home OfficeAnimals Scientific Procedures Act 1986). Bedding was provided on aweekly basis to each cage by use of clean Aspen wood chips (Dates andLtd, Manchester, UK). The bedding was analyzed for specific contaminantsand the results retained on file at Covance. The cages were cleaned anddried before use. Aspen chew blocks were placed within the cages as aform of environmental enrichment. Routinely, holding rooms weremaintained within acceptable limits for temperature and relativehumidity (nominally 19 to 25° C. and 40 to 70%, respectively). Theserooms are illuminated by fluorescent light for 1.2 hours out of each 24hour cycle and designed to receive at least 15 fresh air changes perhour.

RM1.(E).SQC., (Special Diets Services Ltd., Witham, UK) and water fromthe mains tap supply will be provided ad libitum, except where specifiedbelow. These are routinely analyzed for specific constituents and arenot known to contain any biological or chemical entity which mightinterfere with the test system. The treatment groups employed for thestudy were as shown in Table 21: TABLE 21 Treatment Groups. GroupTreatment Dose level (mg/kg) conc. (mg/mL) # of animals 1 Vehicle — — 82 HY-2901  3 0.3 8 3 HY-2901 10 1.0 8 4 HY-2901 30 3.0 8 5 Morphine 100 10.0  8

Measurements of pressure were taken from the left and right hind paws ofeach animal immediately prior to administration of vehicle, test articleor reference compound and at 30, 60, 1.20 and 240 minutes post-oraladministration. The order of the pressure measurements was left pawfollowed by right paw.

Abdominal Spasm Test in the Rat

Each animal received a single administration of vehicle, test article orreference compound by oral gavage, using a constant dose volume 10mg/kg. Individual dose volumes were based on individual body weightsobtained on the day of dosing. The treatment groups are shown in Table22. TABLE 22 Treatment Groups. Group Treatment Dose level (mg/kg) Conc,(mg/mL) # of animals 1 Vehicle — — 6 2 HY-2901  3 0.3 6 3 HY-2901 10 1.06 4 HY-2901 30 3.0 6 5 ASA 100  10.0  6

Forty-five minutes following oral administration each animal received a1 mL intraperitoneal injection of 1% acetic acid. Animals wereimmediately placed into individual observation chambers and the numberof abdominal spasms elicited over the subsequent 25-minute period wasrecorded.

Abdominal Spasm Test in the Mouse

Each animal received a single administration of vehicle, test article orreference compound by oral gavage, using a constant dose volume 10mL/kg. Individual dose volumes were based on individual body weightsobtained on the day of dosing. The treatment groups are shown in Table23. TABLE 23 Treatment Groups. Group Treatment Dose level (mg/kg) Conc.(mg/mL) # of animals 1 Vehicle — — 6 2 HY-2901  3 0.3 6 3 HY-2901 10 1.06 4 HY-2901 30 3.0 6 5 ASA 100  10.0  6

Forty-five minutes following oral administration each animal received a0.25 mL intraperitoneal injection of 0.5% acetic acid. Animals wereimmediately placed into individual observation chambers and the numberof abdominal spasms elicited over the subsequent 25-minute period wasrecorded.

Terminal Procedures

At the end of each test, the animals were humanely killed by a Schedule1 method (e.g. exposure to carbon dioxide gas in a rising concentrationfollowed by dislocation of the neck) and discarded without necropsy. Ifan animal showed any sign of serious discomfort during the study it wassacrificed immediately and humanely. Any animal found dead or killedprematurely during the study was subjected to a necropsy. A macroscopicexamination was performed, after opening the thoracic and abdominalcavities, by observing the appearance of the tissues in situ. Anyabnormalities were recorded.

Results TABLE 24 Effects of HY-2901 on group mean acetic acid-inducedabdominal spasms in the rat following oral administration Dose Orallevel Group mean (=sd) number pf abdominal Group Treatment (mg/kg)spasms recorded in a 25-minute period 1 Vehicle — 33 ± 17.2 (0.25% MC) 2HY-2901  3 31 ± 9.0  3 HY-2901 10 25 ± 16.4 4 HY-2901 30 17 ± 13.8 5 ASA100  1 ± 2.3

TABLE 25 Effects of HY-2901 on acetic acid-induced abdominal spasms inthe rat following oral administration - individual animal data Number ofabdominal Oral Dose level Animal spasms recorded Group Treatment (mg/kg)I.D. in a 25-minute period 1 Vehicle — 1 8 (0.25% MC) 2 41 3 20 4 50 551 6 30 2 HY-2901  3 7 42 8 17 9 26 10 29 11 31 12 39 3 HY-2901 10 13 5714 23 15 20 16 24 17 19 18 9 4 HY-2901 30 19 37 20 20 21 23 22 1 23 1 2418 5 ASA 100  25 0 26 1 27 6 28 1 29 0 30 0

TABLE 26 Effects of HY-2901 on group mean acetic acid-induced abdominalspasms in the mouse following oral administration Dose Oral level Groupmean (±sd) number of abdominal Group Treatment (mg/kg) spasms recordedin a 25 minute period 1 Vehicle — 37 ± 26.8 (0.25% MC) 2 HY-2901  3 6 ±6.2 3 HY-2901 10 1 ± 2.0 4 HY-2901 30 0 ± 0.0 5 ASA 100  29 ± 29.9

TABLE 27 Effects of HY-2901 on acetic acid-induced abdominal spasms inthe mouse following oral administration - individual animal data Numberof abdominal Oral Dose level Animal spasms recorded Group Treatment(mg/kg) I.D. in a 25-minute period 1 Vehicle — 1 2 (0.25% MC) 2 47 3 184 59 5 72 6 23 2 HY-2901  3 7 0 8 1 9 0 10 10 11 7 12 15 3 HY-2901 10 130 14 0 15 0 16 2 17 0 18 5 4 HY-2901 30 19 0 20 0 21 0 22 0 23 0 24 0 5ASA 100  25 17 26 8 27 45 28 82 29 1 30 23

Other Embodiments

While the invention has been described in conjunction with the detaileddescription thereof, the foregoing description is intended to illustrateand not limit the scope of the invention, which is defined by the scopeof the appended claims. Other aspects, advantages, and modifications arewithin the scope of the following claims. It will be understood by thoseskilled in the art that various changes in form and details may be madetherein without departing from the scope of the invention encompassed bythe appended claims.

1. A method of modulating sleep in a subject, comprising administering atherapeutically effective amount of a compound having the formula ofFormula I

or a pharmaceutically effective salt thereof wherein m n, o, p, q are,individually, 0-6, X and Y are, individually, absent, O, S, C(O), SO orSO₂; R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are, independently selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chainalkyl, C₃-C₆ branched alkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃,OCF₃, CH₂OCH₃, CH₂CH₂OCH₃, CH₂OCH₂CH₃, and C₁-C₆ hydroxyalkyl; anyhydrogen in the CH₂ groups in the linker is optionally substituted withH, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chain alkyl, C₃-C₆ branchedalkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃, OCF₃, CH₂OCH₃,CH₂CH₂OCH₃, CH₂OCH₂CH₃, or C₁-C₆ hydroxyalkyl; R₉, R₁₀, R₁₁, and R₁₂are, independently, H, C₁-C₅ straight chain alkyl, C₂-C₆ branched alkyl,R₉ and R₁₀ together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to 7, or R₁₁ and R₁₂ togetherwith the carbon to which they are attached, are connected to form aSpiro ring of size 3 to 7; or substituents on two different atoms areconnected to form a ring of size 3 to 7; R₁₃ and R₁₄ are, independently,selected from the group consisting of H, F, Cl, Br, CH₃, C₁-C₆ straightchain alkyl, and C₂-C₆ branched alkyl; Z is selected from the groupconsisting of CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl,CONHS(O)₂-Heteroaryl, SO₃H, SO₂H, S(O)₂NHCO-alkyl, S(O)₂NHCO-aryl,S(O)NHCO-alkyl, S(O)NHCO-aryl, P(O)(OH)₂, P(O)OH,

further wherein the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 500 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, M3, D1, D2, D3, α1 and α2 that is more than 10 times greaterthan the K_(i) with regard to the H1 receptor; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 13 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 3 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 5 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity relative to thenormal effects of sleep.
 2. The method of claim 1, wherein the compoundhas one or more of the following characteristics: (i) an inhibitionconstant (K_(i)) with regard to H1 receptor binding of less than 150 nM;(ii) a K_(i) with regard to off target binding to an off target selectedfrom the group consisting of M1, M2, and M3, that is greater than 10 μM;(iii) a nonREM peak time value that is greater than 55% nonREM sleep perhour by the third hour after said compound is administered to a subject;(iv) a cumulative total increase in nonREM sleep not less than 20minutes for compound doses that produce maximum sleep consolidation; (v)a longest sleep bout that is greater than 17 minutes in duration; (vi)net longest sleep bout post treatment is greater than or equal to 5minutes when adjusted using a baseline value obtained at least 24 hoursprior to administration of said compound to a subject; (vii) an averagesleep bout that is greater than 6 minutes at absolute peak; (viii)administration of said compound to a subject does not produceappreciable amounts of rebound insomnia; (ix) administration of saidcompound to a subject does not appreciably inhibit REM sleep; and (x)administration of said compound to a subject does not disproportionatelyinhibit locomotor activity or motor tone relative to the normal effectsof sleep.
 3. The method of claim 1, wherein R₉ and R₁₀ and the carbonthey are attached to are absent.
 4. The method of claim 1, wherein R₁₁and R₁₂, together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to
 7. 5. The method of claim 4,wherein R₁₁ and R₁₂ together with the carbon to which they are attached,are connected to form a spiro 3-membered cyclopropyl ring.
 6. The methodof claim 1, wherein Z is CO₂H or


7. The method of claim 1, wherein the compound is selected from thegroup consisting of 47a, 47b, 47c, 47d, 49a, 49b, 49c, 49d, 10a, 10b,10c, 10d, 37, 38a, 38b, 39a and 39b.
 8. The method of claim 1, whereinat least one of R₁-R₈, R₁₃-R₁₄ and at least one of R₉-R₁₀, R₁₁-R₁₂ arenot hydrogen, when Z is COOH.
 9. The method of claim 1, wherein R₁,R₃-R₈ and R₁₃-R₁₄ are each hydrogen and R₂ is not H, CH₃, CF₃, Cl or Br.10. The method of claim 1, wherein R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ are eachhydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br.
 11. Themethod of claim 1, wherein R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ are eachhydrogen, R₆ is CH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br.
 12. Themethod of claim 1, wherein each of R₁, R₃-R₈ and R₁₃-R₁₄ is H and R₂ isF.
 13. The method of claim 1, wherein each of R₁, R₃-R₅, R₇-R₈ andR₁₃-R₁₄ is H, R₆ is OCH₃, and R₂ is OCH₃.
 14. The method of claim 1,wherein each of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ is H, R₆ is F, and R₂ isOCH₃.
 15. The method of claim 1, wherein at least three of R₁-R₈ are nothydrogen.
 16. The method of claim 1, wherein at least one of R₁₃-R₁₄ isnot hydrogen.
 17. The method of claim 1, wherein R₃ is not hydrogen. 18.The method of claim 1, wherein R₇ is not hydrogen.
 19. The method ofclaim 1, wherein R₉ and R₁₀ are each methyl.
 20. The method of claim 1,wherein R₉ and R₁₀ are each ethyl.
 21. The method of claim 1, whereinR₁₁ and R₁₂ are each ethyl.
 22. The method of claim 1, wherein R₁₁ andR₁₂ are each ethyl.
 23. The method of claim 1, wherein R₁₁ and R₁₂ andthe carbon to which they are attached are connected to form a spiro ringof size 3-7.
 24. The method of claim 23, wherein R₁₁ and R₁₂ and thecarbon to which they are attached are connected to form a three-memberedspiro (cyclopropyl) ring.
 25. The method of claim 1, wherein the sleepmodulation is selected from the group consisting of decreasing the timeto sleep onset, increasing the average sleep bout length, and increasingthe maximum sleep bout length.
 26. The method of claim 1 wherein thesleep modulation treats a sleep disorder.
 27. The method of claim 26wherein the sleep disorder is selected from the group consisting ofcircadian rhythm abnormality, insomnia, parasomnia, sleep apneasyndrome, narcolepsy and hypersomnia.
 28. The method of claim 27 whereinthe sleep disorder is circadian rhythm abnormality.
 29. The method ofclaim 27, wherein the sleep disorder is insomnia.
 30. The method ofclaim 27, wherein the sleep disorder is sleep apnea.
 31. The method ofclaim 27, wherein the sleep disorder is narcolepsy.
 32. The method ofclaim 27, wherein the sleep disorder is hypersomnia.
 33. The method ofclaim 1, wherein the compound of Formula I or pharmaceuticallyacceptable salt thereof is administered as a pharmaceutical compositioncomprising a pharmaceutically acceptable excipient.
 34. The method ofclaim 1, wherein the compound of Formula I or pharmaceuticallyacceptable salt thereof is co-administered with one or more additionaltherapies.
 35. The method of claim 1, wherein the subject is selectedfrom the group consisting of humans, companion animals, farm animals,laboratory animals and wild animals.
 36. The method of claim 35, whereinthe subject is a human.
 37. A method of modulating sleep in a subject,comprising administering a therapeutically effective amount of acompound having the formula of Formula II

or a pharmaceutically effective salt thereof wherein m, n, and o are,individually, 0-6, X is absent, O, S, C(O), SO or SO₂; R₁, R₂, R₃, R₆,and R₇ are, independently selected from the group consisting of H, F,Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₃)₂, cyclopropyl, OCH₃, OCF₃, CH₂OCH₃ andCH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅ straight chainalkyl; C₂-C₆ branched alkyl, or R₉ and R₁₀ together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7; R₁₃ and R₁₄ are, independently, selected from the group consisting ofH, F, CH₃, C₁-C₆ straight chain alkyl, and C₂-C₆ branched alkyl, or R₉and R₁₀ together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to 7; Z is selected from thegroup consisting of CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, and

further wherein the compound has one or more of the followingcharacteristics (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 500 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, M3, D1, D2, D3, α1 and α2 that is more than 10 times greaterthan the K_(i) with regard to the H1 receptor; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 13 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 3 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 5 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity relative to thenormal effects of sleep.
 38. The method of claim 37, wherein thecompound has one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 150 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, and M3, that isgreater than 10 μM; (iii) a nonREM peak time value that is greater than55% nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 17minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 5 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 6 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) administration of said compound to a subject does notdisproportionately inhibit locomotor activity or motor tone relative tothe normal effects of sleep.
 39. The method of claim 37, wherein R₉ andR₁₀ and the carbon they are attached to are absent.
 40. The method ofclaim 37, wherein R₉ and R₁₀, together with the carbon to which they areattached, are connected to form a spiro ring of size 3 to
 7. 41. Themethod of claim 40, wherein R₉ and R₁₀, together with the carbon towhich they are attached, are connected to form a spiro cyclopropyl ring.42. The method of claim 37, wherein Z is CO₂H or


43. The method of claim 37, wherein o is zero.
 44. The method of claim37, wherein at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at least one ofR₉-R₁₀, are not hydrogen when Z is COOH.
 45. The method of claim 37,wherein R₁, R₃, R₆-R₇ and R₁₃-R₁₄ are each hydrogen and R₂ is not H,CH₃, CF₃, Cl or Br.
 46. The method of claim 37, wherein R₁, R₃, R₇, R₁₃and R₁₄ are each hydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃, CF₃, Cl orBr.
 47. The method of claim 37, wherein R₁, R₃, R₇, R₁₃ and R₁₄ are eachhydrogen, R₆ is CH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br.
 48. Themethod of claim 37, wherein each of R₁, R₃, R₆, R₇, R₁₃ and R₁₄ is H andR₂ is F.
 49. The method of claim 37, wherein each of R₁, R₃, R₇, R₁₃ andR₁₄ is H, R₆ is OCH₃, and R₂ is OCH₃.
 50. The method of claim 37,wherein each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is F, and R₂ is OCH₃.51. The method of claim 37, wherein at least three of R₁-R₃, R₆-R₇,R₁₃-R₁₄ are not hydrogen.
 52. The method of claim 37, wherein at leastone of R₁₃-R₁₄ is not hydrogen.
 53. The method of claim 37, wherein R₃is not hydrogen.
 54. The method of claim 37, wherein R₇ is not hydrogen.55. The method of claim 37, wherein R₉ and R₁₀ are each methyl.
 56. Themethod of claim 37, wherein R₉ and R₁₀ are each ethyl.
 57. The method ofclaim 37, wherein the sleep modulation is selected from the groupconsisting of decreasing the time to sleep onset, increasing the averagesleep bout length, and increasing the maximum sleep bout length.
 58. Themethod of claim 37 wherein the sleep modulation treats a sleep disorder.59. The method of claim 37, wherein the compound of Formula II orpharmaceutically acceptable salt thereof is administered as apharmaceutical composition comprising a pharmaceutically acceptableexcipient.
 60. The method of claim 37, wherein the compound of FormulaII or pharmaceutically acceptable salt thereof is co-administered withone or more additional therapies.
 61. A method of modulating sleep in asubject, comprising administering a therapeutically effective amount ofa compound having the formula of Formula III

or a pharmaceutically effective salt thereof wherein m and n are,individually, 0-4, X is absent, O, S, C(O), SO or SO₂; R₁ is H, F, Cl,Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, cyclopropyl, CH₂OCH₂CH₃, CH₂OCH₃, CH₂OCH₂CH₃, or OCH₃; R₂, R₃, R₆, and R₇ are, independently, selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, OCH₃,CH₂OCH₃, and CH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅straight chain alkyl; C₂-C₆ branched alkyl, or R₉, and R₁₀, togetherwith the carbon to which they are attached, are connected to form aspiro ring of size 3-7; R₁₃ and R₁₄ are, independently, selected fromthe group consisting of H, F, CH₃, C₁-C₆ straight chain alkyl, C₂-C₆branched alkyl, and CH₂OCH₃; Z is selected from the group consisting ofCO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, and

further wherein the compound has one or more of the followingcharacteristics: (i) an inhibition constant (K_(i)) with regard to H1receptor binding of less than 500 nM; (ii) a K_(i) with regard to offtarget binding to an off target selected from the group consisting ofM1, M2, M3, D1, D2, D3, α1 and α2 that is more than 10 times greaterthan the K_(i) with regard to the H1 receptor; (iii) a nonREM peak timevalue that is greater than 55% nonREM sleep per hour by the third hourafter said compound is administered to a subject; (iv) a cumulativetotal increase in nonREM sleep not less than 20 minutes for compounddoses that produce maximum sleep consolidation; (v) a longest sleep boutthat is greater than 13 minutes in duration; (vi) net longest sleep boutpost treatment is greater than or equal to 3 minutes when adjusted usinga baseline value obtained at least 24 hours prior to administration ofsaid compound to a subject; (vii) an average sleep bout that is greaterthan 5 minutes at absolute peak; (viii) administration of said compoundto a subject does not produce appreciable amounts of rebound insomnia;(ix) administration of said compound to a subject does not appreciablyinhibit REM sleep; and (x) administration of said compound to a subjectdoes not disproportionately inhibit locomotor activity relative to thenormal effects of sleep.
 62. The method of claim 61, wherein thecompound has one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 150 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, and M3, that isgreater than 10 μM; (iii) a nonREM peak time value that is greater than55% nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 17minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 5 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 6 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) administration of said compound to a subject does notdisproportionately inhibit locomotor activity or motor tone relative tothe normal effects of sleep.
 63. The method of claim 61, wherein R₃ isH.
 64. The method of claim 61, wherein R₉ and R₁₀, together with thecarbon to which they are attached, are connected to form a spiro ring ofsize 3-7.
 65. The method of claim 64, wherein R₉ and R₁₀, together withthe carbon to which they are attached, are connected to form a spirocyclopropyl ring.
 66. The method of claim 61, wherein Z is CO₂H or


67. The method of claim 61, wherein at least one of R₁-R₃, R₆-R₇,R₁₃-R₁₄ and at least one of R₉-R₁₀, are not hydrogen when Z is COOH. 68.The method of claim 61, wherein R₁, R₃, R₆-R₇ and R₁₃-R₁₄ are eachhydrogen, and R₂ is not H, CH₃, CF₃, Cl or Br.
 69. The method of claim61, wherein R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂OH, andR₂ is not H, CH₃, CF₃, Cl or Br.
 70. The method of claim 61, wherein R₁,R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂CH₂OH, and R₂ is not H,CH₃, CF₃, Cl or Br.
 71. The method of claim 61, wherein each of R₁, R₃,R₆, R₇, R₁₃ and R₁₄ is H, and R₂ is F.
 72. The method of claim 61,wherein each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is OCH₃, and R₂ isOCH₃.
 73. The method of claim 61, wherein each of R₁, R₃, R₇, R₁₃ andR₁₄ is H, R₆ is F, and R₂ is OCH₃.
 74. The method of claim 61, whereinat least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are not hydrogen.
 75. The methodof claim 61, wherein at least one of R₁₃-R₁₄ is not hydrogen.
 76. Themethod of claim 61, wherein R₃ is not hydrogen.
 77. The method of claim61, wherein R₇ is not hydrogen.
 78. The method of claim 61, wherein R₉and R₁₀ are each methyl.
 79. The method of claim 61, wherein R₉ and R₁₀are each ethyl.
 80. The method of claim 61, wherein the sleep modulationis selected from the group consisting of decreasing the time to sleeponset, increasing the average sleep bout length, and increasing themaximum sleep bout length.
 81. The method of claim 61 wherein the sleepmodulation treats a sleep disorder.
 82. The method of claim 61, whereinthe compound of Formula III or pharmaceutically acceptable salt thereofis administered as a pharmaceutical composition comprising apharmaceutically acceptable excipient.
 83. A method of modulating sleepin a subject, comprising administering a therapeutically effectiveamount of a compound having the formula of Formula IV

or a pharmaceutically effective salt thereof wherein t is 0-6; R₂ and R₆are, independently, H, F, Cl, Br, CF₃, CH₃, OH, OCH₃, CH₂OCH₃, orCH₂OCH₂CH₃; R₉-R₁₀ are H, CH₃ CH₂CH₃, or R₉ and R₁₀, together with thecarbon to which they are attached, are connected to form a spiro ring ofsize 3 to 7; and Z is CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl or

provided that when Z is COOH, t does not equal zero; further wherein thecompound has one or more of the following characteristics: (i) aninhibition constant (K_(i)) with regard to H1 receptor binding of lessthan 500 nM; (ii) a K_(i) with regard to off target binding to an offtarget selected from the group consisting of M1, M2, M3, D1, D2, D3, α1and α2 that is more than 10 times greater than the K_(i) with regard tothe H1 receptor; (iii) a nonREM peak time value that is greater than 55%nonREM sleep per hour by the third hour after said compound isadministered to a subject; (iv) a cumulative total increase in nonREMsleep not less than 20 minutes for compound doses that produce maximumsleep consolidation; (v) a longest sleep bout that is greater than 13minutes in duration; (vi) net longest sleep bout post treatment isgreater than or equal to 3 minutes when adjusted using a baseline valueobtained at least 24 hours prior to administration of said compound to asubject; (vii) an average sleep bout that is greater than 5 minutes atabsolute peak; (viii) administration of said compound to a subject doesnot produce appreciable amounts of rebound insomnia; (ix) administrationof said compound to a subject does not appreciably inhibit REM sleep;and (x) administration of said compound to a subject does notdisproportionately inhibit locomotor activity relative to the normaleffects of sleep.
 84. The method of claim 83, wherein the compound hasone or more of the following characteristics: (i) an inhibition constant(K_(i)) with regard to H1 receptor binding of less than 150 mM; (ii) aK_(i) with regard to off target binding to an off target selected fromthe group consisting of M1, M2, and M3, that is greater than 10 μM;(iii) a nonREM peak time value that is greater than 55% nonREM sleep perhour by the third hour after said compound is administered to a subject;(iv) a cumulative total increase in nonREM sleep not less than 20minutes for compound doses that produce maximum sleep consolidation; (v)a longest sleep bout that is greater than 17 minutes in duration; (vi)net longest sleep bout post treatment is greater than or equal to 5minutes when adjusted using a baseline value obtained at least 24 hoursprior to administration of said compound to a subject; (vii) an averagesleep bout that is greater than 6 minutes at absolute peak; (viii)administration of said compound to a subject does not produceappreciable amounts of rebound insomnia; (ix) administration of saidcompound to a subject does not appreciably inhibit REM sleep; and (x)administration of said compound to a subject does not disproportionatelyinhibit locomotor activity or motor tone relative to the normal effectsof sleep.
 85. The method of claim 83, wherein R₉ and R₁₀ together withthe carbon to which they are attached, are connected to form a spiroring of size 3 to
 7. 86. The method of claim 85, wherein R₉ and R₁₀together with the carbon to which they are attached, are connected toform a spiro 3-membered cyclopropyl ring.
 87. The method of claim 83,wherein Z is CO₂H or


88. The method of claim 83, wherein at least one of R₂, and R₆ and atleast one of R₉-R₁₀, are not hydrogen when Z is COOH.
 89. The method ofclaim 83, wherein R₆ is hydrogen, and R₂ is not H, CH₃, CF₃, Cl or Br.90. The method of claim 83, wherein R₆ is CH₂OH, and R₂ is not H, CH₃,CF₃, Cl or Br.
 91. The method of claim 83, wherein R₆ is CH₂CH₂OH, andR₂ is not H, CH₃, CF₃, Cl or Br.
 92. The method of claim 83, wherein R₆is H, and R₂ is F.
 93. The method of claim 83, wherein R₆ is OCH₃, andR₂ is OCH₃.
 94. The method of claim 83, wherein R₆ is F, and R₂ is OCH₃.95. The method of claim 83, wherein R₉ and R₁₀ are each methyl.
 96. Themethod of claim 83, wherein R₉ and R₁₀ are each ethyl.
 97. The method ofclaim 83, wherein the sleep modulation is selected from the groupconsisting of decreasing the time to sleep onset, increasing the averagesleep bout length, and increasing the maximum sleep bout length.
 98. Themethod of claim 83 wherein the sleep modulation treats a sleep disorder.99. The method of claim 83, wherein the compound of Formula IV orpharmaceutically acceptable salt thereof is administered as apharmaceutical composition comprising a pharmaceutically acceptableexcipient.
 100. The method of claim 83, wherein the compound of FormulaI or pharmaceutically acceptable salt thereof is co-administered withone or more additional therapies.
 101. The method of claim 83, whereinthe compound is selected from formula IVa, IVb, IVc, and IVd.
 102. Amethod of modulating sleep in a subject, comprising administering atherapeutically effective amount of compound 10a:


103. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 47a:


104. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 49a:


105. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 49b:


106. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 37:


107. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 38a:


108. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 38b:


109. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 39a:


110. A method of modulating sleep in a subject, comprising administeringa therapeutically effective amount of compound 39b:


111. A compound according to Formula I

or a pharmaceutically effective salt thereof wherein m n, o, p, q are,individually, 0-6, X and Y are, individually, absent, O, S, C(O), SO orSO₂; R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are, independently selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chainalkyl, C₃-C₆ branched alkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃,OCF₃, CH₂OCH₃, CH₂CH₂OCH₃, CH₂OCH₂CH₃, and C₁-C₆ hydroxyalkyl; anyhydrogen in the CH₂ groups in the linker is optionally substituted withH, F, Cl, Br, CF₃, CH₃, C₂-C₆ straight chain alkyl, C₃-C₆ branchedalkyl, C₃-C₇ cycloalkyl, C₃-C₇ heterocyclyl, OCH₃, OCF₃, CH₂OCH₃,CH₂CH₂OCH₃, CH₂OCH₂CH₃, or C₁-C₆ hydroxyalkyl; R₉, R₁₀, R₁₁, and R₁₂are, independently, H, C₁-C₅ straight chain alkyl, C₂-C₆ branched alkyl,R₉ and R₁₀ together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to 7, or R₁₁ and R₁₂ togetherwith the carbon to which they are attached, are connected to form aspiro ring of size 3 to 7; or substituents on two different atoms areconnected to form a ring of size 3 to 7; R₁₃ and R₁₄ are, independently,selected from the group consisting of H, F, Cl, Br, CH₃, C₁-C₆ straightchain alkyl, and C₂-C₆ branched alkyl; Z is selected from the groupconsisting of CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl,CONHS(O)₂-Heteroaryl, SO₃H, SO₂H, S(O)₂NHCO-alkyl, S(O)₂NHCO-aryl,S(O)NHCO-alkyl, S(O)NHCO-aryl, P(O)(OH)₂, P(O)OH,


112. The compound of claim 111, wherein R₉ and R₁₀ and the carbon theyare attached to are absent.
 113. The compound of claim 111, wherein R₁₁and R₁₂, together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to
 7. 114. The compound ofclaim 111, wherein R₁₁ and R₁₂ together with the carbon to which theyare attached, are connected to form a spiro 3-membered cyclopropyl ring.115. The compound of claim 111, wherein Z is CO₂H or


116. The compound of claim 111, wherein the compound is selected fromthe group consisting of 47a, 47b, 47c, 47d, 49a, 49b, 49c, 49d, 10a,10b, 10c, 10d, 37, 38a, 38b, 39a and 39b.
 117. The compound of claim111, wherein at least one of R₁-R₈, R₁₃-R₁₄ and at least one of R₉-R₁₀,R₁₁-R₁₂ are not hydrogen, when Z is COOH.
 118. The compound of claim111, wherein R₁, R₃-R₈ and R₁₃-R₁₄ are each hydrogen and R₂ is not H,CH₃, CF₃, Cl or Br.
 119. The compound of claim 111, wherein R₁, R₃-R₅,R₇-R₈ and R₁₃-R₁₄ are each hydrogen, R₆ is CH₂OH, and R₂ is not H, CH₃,CF₃, Cl or Br.
 120. The compound of claim 111, wherein R₁, R₃-R₅, R₇-R₈and R₁₃-R₁₄ are each hydrogen, R₆ is CH₂CH₂OH, and R₂ is not H, CH₃,CF₃, Cl or Br.
 121. The compound of claim 111, wherein each of R₁, R₃-R₈and R₁₃-R₁₄ is H and R₂ is F.
 122. The compound of claim 111, whereineach of R₁, R₃-R₅, R₇-R₈ and R₁₃-R₁₄ is H, R₆ is OCH₃, and R₂ is OCH₃.123. The compound of claim 111, wherein each of R₁, R₃-R₅, R₇-R₈ andR₁₃-R₁₄ is H, R₆ is F, and R₂ is OCH₃.
 124. The compound of claim 111,wherein at least three of R₁-R₈ are not hydrogen.
 125. The compound ofclaim 111, wherein at least one of R₁₃-R₁₄ is not hydrogen.
 126. Themethod of claim 1, wherein at least one of R₃ and R₇ is not hydrogen.127. The compound of claim 111, wherein R₉ and R₁₀ are each methyl. 128.The compound of claim 111, wherein R₉ and R₁₀ are each ethyl.
 129. Thecompound of claim 111, wherein R₁₁ and R₁₂ are each ethyl.
 130. Thecompound of claim 111, wherein R₁₁ and R₁₂ are each ethyl.
 131. Thecompound of claim 111, wherein R₁₁ and R₁₂ and the carbon to which theyare attached are connected to form a spiro ring of size 3-7.
 132. Thecompound of claim 131, wherein R₁₁ and R₁₂ and the carbon to which theyare attached are connected to form a three-membered spiro (cyclopropyl)ring.
 133. The compound of claim 111, further comprising apharmaceutically acceptable excipient.
 134. A compound of Formula II

or a pharmaceutically effective salt thereof wherein m, n, and o are,individually, 0-6, X is absent, O, S, C(O), SO or SO₂; R₁, R₂, R₃, R₆,and R₇ are, independently selected from the group consisting of H, F,Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₃)₂, cyclopropyl, OCH₃, OCF₃, CH₂OCH₃ andCH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅ straight chainalkyl; C₂-C₆ branched alkyl, or R₉ and R₁₀ together with the carbon towhich they are attached, are connected to form a spiro ring of size 3 to7; R₁₃ and R₁₄ are, independently, selected from the group consisting ofH, F, CH₃, C₁-C₆ straight chain alkyl, and C₂-C₆ branched alkyl, or R₉and R₁₀ together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to 7; Z is selected from thegroup consisting of CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, and


135. The compound of claim 134, wherein R₉ and R₁₀ and the carbon theyare attached to are absent.
 136. The compound of claim 134, wherein R₉and R₁₀, together with the carbon to which they are attached, areconnected to form a spiro ring of size 3 to
 7. 137. The compound ofclaim 136, wherein R₉ and R₁₀, together with the carbon to which theyare attached, are connected to form a spiro cyclopropyl ring.
 138. Thecompound of claim 134, wherein Z is CO₂H or


139. The compound of claim 134, wherein o is zero.
 140. The compound ofclaim 134, wherein at least one of R₁-R₃, R₆-R₇, R₁₃-R₁₄ and at leastone of R₉-R₁₀, are not hydrogen when Z is COOH.
 141. The compound ofclaim 134, wherein R₁, R₃, R₆-R₇ and R₁₃-R₁₄ are each hydrogen and R₂ isnot H, CH₃, CF₃, Cl or Br.
 142. The compound of claim 134, wherein R₁,R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂OH, and R₂ is not H,CH₃, CF₃, Cl or Br.
 143. The compound of claim 134, wherein R₁, R₃, R₇,R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂CH₂OH, and R₂ is not H, CH₃,CF₃, Cl or Br.
 144. The compound of claim 134, wherein each of R₁, R₃,R₆, R₇, R₁₃ and R₁₄ is H and R₂ is F.
 145. The compound of claim 134,wherein each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is OCH₃, and R₂ isOCH₃.
 146. The compound of claim 134, wherein each of R₁, R₃, R₇, R₁₃and R₁₄ is H, R₆ is F, and R₂ is OCH₃.
 147. The compound of claim 134,wherein at least one of R₁₃-R₁₄ is not hydrogen.
 148. The compound ofclaim 134, wherein R₉ and R₁₀ are each methyl.
 149. The compound ofclaim 134, wherein R₉ and R₁₀ are each ethyl.
 150. A compound of FormulaIII

or a pharmaceutically effective salt thereof wherein m and n are,individually, 0-4, X is absent, O, S, C(O), SO or SO₂; R₁ is H, F, Cl,Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, cyclopropyl, CH₂OCH₂CH₃, CH₂OCH₃, CH₂OCH₂CH₃, or OCH₃; R₂, R₃, R₆, and R₇ are, independently, selected fromthe group consisting of H, F, Cl, Br, CF₃, CH₃, CH₂CH₃, CH(CH₂)₂, OCH₃,CH₂OCH₃, and CH₂OCH₂CH₃; R₉, and R₁₀, are, independently, H, C₁-C₅straight chain alkyl; C₂-C₆ branched alkyl, or R₉, and R₁₀, togetherwith the carbon to which they are attached, are connected to form aSpiro ring of size 3-7; R₁₃ and R₁₄ are, independently, selected fromthe group consisting of H, F, CH₃, C₁-C₆ straight chain alkyl, C₂-C₆branched alkyl, and CH₂OCH₃; Z is selected from the group consisting ofCO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl, and


151. The compound of claim 150, wherein R₃ is H.
 152. The compound ofclaim 150, wherein R₉ and R₁₀, together with the carbon to which theyare attached, are connected to form a spiro ring of size 3-7.
 153. Thecompound of claim 152, wherein R₉ and R₁₀, together with the carbon towhich they are attached, are connected to form a spiro cyclopropyl ring.154. The compound of claim 150, wherein Z is CO₂H or


155. The compound of claim 150, wherein at least one of R₁-R₃, R₆-R₇,R₁₃-R₁₄ and at least one of R₉-R₁₀, are not hydrogen when Z is COOH.156. The compound of claim 150, wherein R₁, R₃, R₆-R₇ and R₁₃-R₁₄ areeach hydrogen, and R₂ is not H, CH₃, CF₃, Cl or Br.
 157. The compound ofclaim 150, wherein R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ isCH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br.
 158. The compound of claim150, wherein R₁, R₃, R₇, R₁₃ and R₁₄ are each hydrogen, R₆ is CH₂CH₂OH,and R₂ is not H, CH₃, CF₃, Cl or Br.
 159. The compound of claim 150,wherein each of R₁, R₃, R₆, R₇, R₁₃ and R₁₄ is H, and R₂ is F.
 160. Thecompound of claim 150, wherein each of R₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆is OCH₃, and R₂ is OCH₃.
 161. The compound of claim 150, wherein each ofR₁, R₃, R₇, R₁₃ and R₁₄ is H, R₆ is F, and R₂ is OCH₃.
 162. The compoundof claim 150, wherein at least three of R₁-R₃, R₆-R₇, R₁₃-R₁₄ are nothydrogen.
 163. The compound of claim 150, wherein at least one ofR₁₃-R₁₄ is not hydrogen.
 164. The compound of claim 150, wherein R₉ andR₁₀ are each methyl.
 165. The compound of claim 150, wherein R₉ and R₁₀are each ethyl.
 166. A compound of Formula IV

or a pharmaceutically effective salt thereof wherein t is 0-6; R₂ and R₆are, independently, H, F, Cl, Br, CF₃, CH₃, OH, OCH₃, CH₂OCH₃, orCH₂OCH₂CH₃; R₉-R₁₀ are H, CH₃ CH₂CH₃, or R₉ and R₁₀, together with thecarbon to which they are attached, are connected to form a Spiro ring ofsize 3 to 7; and Z is CO₂H, CONHS(O)₂-Aryl, CONHS(O)₂-Alkyl or

provided that when Z is COOH, t does not equal zero.
 167. The compoundof claim 166, wherein R₉ and R₁₀ together with the carbon to which theyare attached, are connected to form a spiro ring of size 3 to
 7. 168.The compound of claim 167, wherein R₉ and R₁₀ together with the carbonto which they are attached, are connected to form a spiro 3-memberedcyclopropyl ring.
 169. The compound of claim 166 wherein Z is CO₂H or


170. The compound of claim 166, wherein at least one of R₂, and R₆ andat least one of R₉-R₁₀, are not hydrogen when Z is COOH.
 171. Thecompound of claim 166, wherein R₆ is hydrogen, and R₂ is not H, CH₃,CF₃, Cl or Br.
 172. The compound of claim 166, wherein R₆ is CH₂OH, andR₂ is not H, CH₃, CF₃, Cl or Br.
 173. The compound of claim 166, whereinR₆ is CH₂CH₂OH, and R₂ is not H, CH₃, CF₃, Cl or Br.
 174. The compoundof claim 166, wherein R₉ and R₁₀ are each methyl.
 175. The compound ofclaim 166, wherein R₉ and R₁₀ are each ethyl.
 176. A compound having thestructure of compound 10a:


177. A compound having the structure of compound 47a:


178. A compound having the structure of compound 49a:


179. A compound having the structure of compound 49b:


180. A compound having the structure of compound 37:


181. A compound having the structure of compound 38a:


182. A compound having the structure of compound 38b:


183. A compound having the structure of compound 39a:


184. A compound having the structure of compound 39b: